Anti-cancer nuclear hormone receptor-targeting compounds

ABSTRACT

The disclosure relates to anti-cancer compounds derived from nuclear steroid receptor binders, to products containing the same, as well as to methods of their use and preparation.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 3.5 U.S.C. § 119(e) of U.S.Provisional Application 62/671,382, filed on May 14, 2018, which ishereby incorporated by reference in its entirety.

BACKGROUND

The disclosure relates to anti-cancer compounds derived from nuclearreceptor binders, such as nuclear steroid receptor binders, to productscontaining the same, as well as to methods of their use and preparation.

PARP inhibitors are pharmacologic agents that prevent DNA repair leadingto the death of cells and hence tumor growth inhibition. This mechanismof preventing cell growth leads to significant anti tumor activity intumors with BRCA1, BRCA2 and PALB2 mutations, as these proteins areimportant for the repair of double strand DNA breaks by the homologousrecombinant repair (HRR) pathway. Normal cells that are not dividing asfast as tumors and do not carry mutated BRCA1 or BRCA2 still have theHRR pathway intact which allows them to survive in the face of PARPinhibition. In addition to the catalytic inhibition of PARP, researchersat the National Cancer Institute in 2012 discovered an additionalmechanism that drives the toxic effect of PARP inhibitors in tumorcells. Their observation that in addition to the blockade of theenzymatic activity of PARP, certain PARP inhibitors have the ability tolocalize PARP proteins to sites of DNA damage, correlated with thecytotoxicity of these inhibitors. This mode of action, called “PARPtrapping” is an additional mechanism by which this class ofpharmacologic agents works in the preventing tumor growth and survival(Murai, et al. Cancer Research (2012) 72(21): 5588-99). Inhibitors ofPARP enzymes (such as olaparib, rucaparib, niraparib, and talzoparib)have been approved for the treatment of breast cancer in patients withBRCA mutations, and ovarian cancer. There are several others (e.g.,velaparib) that are in clinical testing for breast, prostate and ovariancancers. The use of PARP inhibitors is not without side effects, and oneof the major road blocks to the long-term use of PARP inhibitors is therapid and dose dependent development of neutropenia. This requiresdosing holidays and/or dose reductions in clinical practice, whichcompromise the ability to achieve maximal efficacy.

SUMMARY

Provided herein are compounds comprising at least one nuclear payloadand at least one nuclear receptor-targeting epitope. Compounds describedherein are designed to bind nuclear receptors within the cell and allowthe compound, with its nuclear payload, to accumulate in the nucleus.

Also provided herein are compounds comprising at least one nuclearpayload and at least one nuclear steroid receptor-targeting epitope.Compounds described herein are designed to bind nuclear steroidreceptors within the cell (e.g., the androgen receptor in the cytosol)and allow the compound, with its nuclear payload, to accumulate in thenucleus. This approach allows for compounds to have cell-typeselectivity, not merely improved potency, and possibly a highertherapeutic index.

Further, the compounds described herein offer targeted delivery of anuclear payload. The compounds both target and localize within tumortissue. The transport of the compound, which comprises at least onenuclear receptor-targeting epitope, such as a nuclear steroidreceptor-targeting epitope, covalently attached to at least one nuclearpayload, to the nucleus allows for accumulation of the nuclear payloadin the nucleus, enhancing tumor cell death. By doing so, compoundsdescribed in this disclosure may exhibit superior efficacy. In addition,the compounds described in this disclosure will, by accumulatingpreferentially in the of nuclear receptor positive cells, such assteroid receptor positive cells, spare cells that do not express thespecific nuclear steroid receptor, and therefore reduce side effects.

In certain embodiments, provided is a compound comprising at least onenuclear payload and at least one nuclear receptor-targeting epitope. Incertain embodiments, when the compound comprises one nuclear payload andone nuclear receptor-targeting epitope, the nuclear receptor-targetingepitope is not a peptide, protein, nanoparticle or antibody. In certainembodiments, when the compound comprises one nuclear payload and onenuclear receptor-targeting epitope, where the nuclear receptor-targetingepitope is an androgen receptor-targeting epitope or an estrogenreceptor-targeting epitope, the nuclear payload is not doxorubicin, oran analog thereof. In certain embodiments, when the compound comprisesone nuclear payload and one nuclear receptor-targeting epitope, wherethe nuclear receptor-targeting epitope is an androgen receptor-targetingepitope or an estrogen receptor-targeting epitope, the nuclear payloadis not a hydroxamic acid which binds histone deacetylase (HDAC).

In certain embodiments, provided is a compound comprising at least onenuclear payload and at least one nuclear receptor-targeting epitope;provided that when the compound comprises one nuclear payload and onenuclear receptor-targeting epitope:

the nuclear receptor-targeting epitope is not a peptide, protein,nanoparticle or antibody; and

when the nuclear receptor-targeting epitope is an androgenreceptor-targeting epitope or an estrogen receptor-targeting epitope,the nuclear payload is not doxorubicin, or an analog thereof, and is nota hydroxamic acid which binds histone deacetylase (HDAC).

In certain embodiments, provided is a compound comprising at least onenuclear payload which binds to a catalytic domain of poly(ADP-ribose)polymerase (PARP) and at least one nuclear receptor-targeting epitope.In certain embodiments, the at least one nuclear receptor-targetingepitope is a nuclear steroid receptor-targeting epitope.

In certain embodiments, provided is a compound comprising at least onenuclear payload which binds to a poly(ADP-ribose) polymerase (PARP)(e.g., PARP-1 and/or PARP-2) and at least one nuclear receptor-targetingepitope. In certain embodiments, provided is a compound comprising atleast one nuclear payload which binds to a poly(ADP-ribose) polymerase(PARP) (e.g., PARP-1 and/or PARP-2) and at least one estrogenreceptor-targeting epitope, glucocorticoid receptor-targeting epitope,progesterone receptor-targeting epitope or androgen receptor-targetingepitope.

In certain embodiments, provided is a compound comprising at least onenuclear payload which binds to poly(ADP-ribose) polymerase (PARP) (e.g.,PARP-1 and/or PARP-2) and at least one nuclear receptor-targetingepitope derived from an androgen receptor agonist, an androgen receptorantagonist, a selective androgen-receptor modulator (SARM), an estrogenreceptor agonist, an estrogen receptor antagonist, a selective estrogenreceptor modulator (SERM), a selective estrogen receptor modulator(SERM), a glucocorticoid receptor agonist, a glucocorticoid receptorantagonist, a selective glucocorticoid receptor modulator (SGRM), aprogesterone receptor antagonist, a progesterone receptor agonist, aselective progesterone receptor modulator (SPRM), or a combinationthereof.

In certain embodiments, provided is a compound comprising at least onenuclear payload which binds to poly(ADP-ribose) polymerase (PARP) (e.g.,PARP-1 and/or PARP-2) and at least one nuclear steroidreceptor-targeting epitope derived from an androgen receptor agonist, anandrogen receptor antagonist, a selective androgen-receptor modulator(SARM), an estrogen receptor agonist, an estrogen receptor antagonist, aprogesterone receptor antagonist, a progesterone receptor agonist, aselective progesterone receptor modulator (SPRM), or a combinationthereof.

In certain embodiments, provided is a compound comprising at least onenuclear payload which binds to a poly(ADP-ribose) polymerase (PARP)(e.g., PARP-1 and/or PARP-2) and at least one nuclear receptor-targetingepitope derived from estrogen, estetrol, estriol, estrone, progesterone,enobosarm, bicalutamide, apalutamide, testosterone, dihydrotestosterone,estradiol, flutamide, nilutamide, enzalutamide, tamoxifen, toremifene,raloxifene, bazedoxifene, ospemifene, megestrol acetate, estramustine,abiraterone, LGD-2941, BMS-564929, ostarine, or an analog thereof.

Also provided is a compound of Formula I or II, or a stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

A-(L-B)_(m)  I

A-L-(B)_(m)  II

wherein:

A is a nuclear payload;

m is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope; and

each L is independently a covalent bond or a linking moiety.

In certain embodiments of Formula I or II, when m is 1, the nuclearreceptor-targeting epitope is not a peptide, protein, nanoparticle orantibody. In certain embodiments of Formula I or II, when m is 1, and Bis an androgen receptor-targeting epitope or an estrogenreceptor-targeting epitope, then A is not doxorubicin, or an analogthereof. In certain embodiments of Formula I or II, when m is 1, and Bis an androgen receptor-targeting epitope or an estrogenreceptor-targeting epitope, then A is not a hydroxamic acid which bindshistone deacetylase (HDAC).

Also provided is compound of Formula III, IV, V or VI, or stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

wherein:

each R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more (e.g., 1 to 5, or 1 to 3) R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more (e.g., 1 to 5, or 1 to 3) haloor C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino asvalency permits;

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino;

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino; and

R⁹ is hydrogen or R²;

provided that at least one R¹, R², R³ and R⁴ is -L-(B)_(m).

Also provided is compound of Formula IIIA, IV, VA or VI, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more (e.g., 1 to 5, or 1 to 3) R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl,

-   -   C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, aryl, heteroaryl,        —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷, —OC(═O)R⁷, —C(═O)NR⁷R⁸,        —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷, —S(═O)₁₋₂NR⁷R⁸,        —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸, —NR⁷C(═O)OR⁸ or        —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl, cycloalkyl,        heterocyclyl, aryl and heteroaryl of R¹⁰ are independently        optionally substituted with one or more (e.g., 1 to 5, or 1        to 3) halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo,        hydroxyl or amino as valency permits;

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

Also provided is compound of Formula IIIB, IVA, VB or VIA, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

Also provided is a compound of Table 1, or stereoisomer, mixture ofstereoisomers, hydrate, solvate, isotopically enriched analog orpharmaceutically acceptable salt thereof.

Also provided is a composition comprising a compound as described hereinor stereoisomer, mixture of stereoisomers, hydrate, solvate,isotopically enriched analog or pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable excipient.

Also provided is a method of treating or preventing cancer, comprisingadministering an effective amount of a compound or composition asdescribed herein to an individual in need thereof. The cancer can be ablood cancer, lung cancer, breast cancer, fallopian tube cancer, braincancer, head and neck cancer, esophageal cancer, ovarian cancer,pancreatic cancer, peritoneal cancer, prostate cancer or skin cancer,such as, but not limited to, liver cancer, melanoma, Hodgkin's disease,non-Hodgkin's lymphomas, acute lymphocytic leukemia, chronic lymphocyticleukemia, multiple myeloma, neuroblastoma, breast carcinoma, ovariancarcinoma, lung carcinoma, Wilms' tumor, cervical carcinoma, testicularcarcinoma, soft-tissue sarcoma, chronic lymphocytic leukemia, primarymacroglobulinemia, bladder carcinoma, chronic granulocytic leukemia,primary brain carcinoma, malignant melanoma, small-cell lung carcinoma,stomach carcinoma, colon carcinoma, malignant pancreatic insulinoma,malignant carcinoid carcinoma, malignant melanoma, choriocarcinoma,mycosis fungoide, head neck carcinoma, osteogenic sarcoma, pancreaticcarcinoma, acute granulocytic leukemia, hairy cell leukemia,rhabdomyosarcoma, Kaposi's sarcoma, genitourinary carcinoma, thyroidcarcinoma, esophageal carcinoma, malignant hypercalcemia, cervicalhyperplasia, renal cell carcinoma, endometrial carcinoma, polycythemiavera, essential thrombocytosis, adrenal cortex carcinoma, skin cancer,or prostatic carcinoma.

Also provided is a method of treating or preventing bladder cancer,breast cancer, fallopian tube cancer, ovarian cancer, prostate cancer,peritoneal cancer, testicular cancer, endometrial cancer, or uterinecancer, comprising administering an effective amount of a compound orcomposition as described herein, or a pharmaceutically acceptable saltor solvate thereof, to an individual in need thereof.

Also provided is a method of treating or preventing cancer, comprisingadministering an effective amount of a compound, or a pharmaceuticallyacceptable salt or solvate thereof, comprising at least one nuclearpayload and at least one nuclear receptor-targeting epitope comprisingtestosterone, a testosterone ester (e.g., testosterone enanthate,propionate, cypionate, etc., or an analog thereof), enobosarm,BMS-564929, PS178990, LGD-4033 (ligandrol), LGD-2941, AC-262,356,JNJ-28330835, JNJ-37654032, JNJ-26146900, LGD-2226, LGD-3303,LGD-121071, LG-120907, S-40503, S-23, RAD-140, acctothiolutamide,andarinc (S-4), LG-121071, TFM-4AS-1, YK-11, MK-0773 (PF-05314882),GSK2849466, GSK2881078, GSK8698, GSK4336, ACP-105, TT701, LY2452473, a1-(2-hydroxy-2-methyl-3-phenoxypropanoyl)-indoline-4-carbonitrile-derivative(J Med Chem. 2014, 57(6), 2462-71), anordrin, bazedoxifene, broparestrol(Acnestrol), clomifene (Clomid), cyclofenil (Sexovid), lasofoxifene(Fablyn), ormeloxifene (Centron, Novex, Novex-DS, Sevista), ospemifene(Osphena, deaminohydroxytoremifene), raloxifene (Evista), tamoxifen(Nolvadex), toremifene (Fareston; 4-chlorotamoxifen), acolbifene,afimoxifene (4-hydroxytamoxifen; metabolite of tamoxifen), elacestrant,enclomifene ((E)-clomifene), endoxifen (4-hydroxy-N-desmethyltamoxifen;metabolite of tamoxifen), zuclomifene ((Z)-clomifene), bazedoifene,arzoxifene, brilanestrant, clomifenoxide (clomiphene N-oxide; metaboliteof clomifene), droloxifene (3-hydroxytamoxifen), etacstil, fispemifene,GW-7604 (4-hydroxyetacstil), idoxifene (pyrrolidino-4-iodotamoxifen),levormeloxifene ((L)-ormeloxifene), miproxifene, nafoxidine, nitromifene(CI-628), panomifene, pipendoxifene (ERA-923), trioxifene, keoxifene,LY117018, onapristone, fareston (toremifine citrate), zindoxifene(D-16726), fulvestrant, ARN-810, GW5638, GW7604, ulipristal acetate,asoprisnil (J867), mifepristone, telapristone (CDB-4124, Proellex,Progenta), or an analog thereof. In certain embodiments, the nuclearpayload comprises a PARP inhibitor.

Also provided is a method of treating or preventing an androgen receptoroverexpressing cancer, comprising administering an effective amount of acompound, or a pharmaceutically acceptable salt or solvate thereof,comprising at least one nuclear payload and at least one androgenreceptor-targeting epitope to an individual in need thereof. In certainembodiments, the cancer is prostate, breast, triple negative breastcancer, bladder, or liver cancer. In certain embodiments, the androgenreceptor-targeting epitope comprises an androgen receptor agonist, aselective androgen-receptor modulator (SARM), an androgen receptorantagonist, a selective estrogen receptor modulator (SERM), an estrogenreceptor antagonist, a progestin, or an estrogen. In certainembodiments, the androgen receptor-targeting epitope comprisesenobosarm, bicalutamide, flutamide, nilutamide, enzalutamide, tamoxifen,toremifene, raloxifene, fulvestrant, megestrol acetate, estramustine,ketoconazole, abiraterone, darolutamide, or an analog thereof. Incertain embodiments, the androgen receptor-targeting epitope comprisesenobosarm, bicalutamide, flutamide, nilutamide, enzalutamide, tamoxifen,toremifene, raloxifene, fulvestrant, megestrol acetate, estramustine,ketoconazole, abiraterone, or an analog thereof. In certain embodiments,the nuclear payload comprises a PARP inhibitor.

Also provided is a method of treating or preventing an estrogen and/orprogesterone receptor overexpressing cancer, comprising administering aneffective amount of a compound, or a pharmaceutically acceptable salt orsolvate thereof, comprising at least one nuclear payload and at leastone estrogen and/or progesterone receptor-targeting epitope to anindividual in need thereof. In certain embodiments, the cancer isbreast, uterine, or ovarian cancer. In certain embodiments, the nuclearpayload comprises a PARP inhibitor.

Also provided is a method of treating or preventing a glucocorticoidreceptor overexpressing cancer, comprising administering an effectiveamount of a compound, or a pharmaceutically acceptable salt or solvatethereof, comprising at least one nuclear payload and at least oneglucocorticoid receptor-targeting epitope to an individual in needthereof. In certain embodiments, the cancer is prostate, breast,uterine, or ovarian cancer. In certain embodiments, the nuclear payloadcomprises a PARP inhibitor.

Also provided is a method of treating or preventing cancer, comprisingadministering an effective amount of a compound or composition asdescribed herein, or a pharmaceutically acceptable salt or solvatethereof, in combination with an additional chemotherapeutic agent, to anindividual in need thereof.

Also provided is a method for the treatment or prevention of a conditionwhich can be ameliorated by inhibition of PARP in an individual in needthereof, the method comprising administering to the individual aneffective amount of a compound or composition of any preceding claim ora pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION

The following description sets forth exemplary embodiments of thepresent technology. It should be recognized, however, that suchdescription is not intended as a limitation on the scope of the presentdisclosure but is instead provided as a description of exemplaryembodiments.

1. Definitions

As used in the present specification, the following words, phrases andsymbols are generally intended to have the meanings as set forth below,except to the extent that the context in which they are used indicatesotherwise.

The term “about” refers to a variation of ±1%, ±3%, ±5%, or ±10% of thevalue specified. For example, “about 50” can in some embodimentsincludes a range of from 45 to 55. For integer ranges, the term “about”can include one or two integers greater than and/or less than a recitedinteger at each end of the range. Unless indicated otherwise herein, theterm “about” is intended to include values, e.g., weight percentages,proximate to the recited range that are equivalent in terms of thefunctionality of the individual ingredient, the composition, or theembodiment. Also, the singular forms “a” and “the” include pluralreferences unless the context clearly dictates otherwise. Thus, e.g.,reference to “the compound” includes a plurality of such compounds andreference to “the assay” includes reference to one or more compounds andequivalents thereof known to those skilled in the art.

“Alkyl” refers to an unbranched or branched saturated hydrocarbon chain.As used herein, alkyl has 1 to 10 carbon atoms (i.e., C₁₋₁₀ alkyl), 1 to8 carbon atoms (i.e., C₁₋₈ alkyl), 1 to 6 carbon atoms (i.e., C₁₋₆alkyl), or 1 to 4 carbon atoms (i.e., C₁₋₄ alkyl). Examples of alkylgroups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl,iso-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl,2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl residue having aspecific number of carbons is named by chemical name or identified bymolecular formula, all positional isomers having that number of carbonsmay be encompassed; thus, for example, “butyl” includes n-butyl (i.e.—(CH₂)₃CH₃), sec-butyl (i.e. —CH(CH₃)CH₂CH₃), isobutyl (i.e.—CH₂CH(CH₃)₂) and tert-butyl (i.e. —C(CH₃)₃); and “propyl” includesn-propyl (i.e. —(CH₂)₂CH₃) and isopropyl (i.e. —CH(CH₃)₂).

“Haloalkyl” refers to an unbranched or branched alkyl group as definedabove, wherein one or more hydrogen atoms are replaced by a halogen. Forexample, where a residue is substituted with more than one halogen, itmay be referred to by using a prefix corresponding to the number ofhalogen moieties attached. Dihaloalkyl and trihaloalkyl refer to alkylsubstituted with two (“di”) or three (“tri”) halo groups, which may be,but are not necessarily, the same halogen. Examples of haloalkyl includedifluoromethyl (—CHF₂) and trifluoromethyl (—CF₃).

“Heteroalkyl” refers to an alkyl group in which one or more of thecarbon atoms (and any associated hydrogen atoms) are each independentlyreplaced with the same or different heteroatomic group. The term“heteroalkyl” includes unbranched or branched saturated chain havingcarbon and heteroatoms. By way of example, 1, 2 or 3 carbon atoms may beindependently replaced with the same or different heteroatomic group.Heteroatomic groups include, but are not limited to, —NH—, —O—, —S—,—S(O)—, —S(O)₂—, and the like. As used herein, heteroalkyl includes 1 to8 carbon atoms, or 1 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2heteroatoms, or 1 heteroatom.

“Alkoxy” refers to the group “—O-alkyl”.

“Alkenyl” refers to an alkyl group containing at least one carbon-carbondouble bond and having from 2 to 20 carbon atoms (i.e., C₂₋₂₀ alkenyl),2 to 8 carbon atoms (i.e., C₂₋₈ alkenyl), 2 to 6 carbon atoms (i.e.,C₂₋₆ alkenyl) or 2 to 4 carbon atoms (i.e., C₂₋₄ alkenyl). Examples ofalkenyl groups include, e.g., ethenyl, propenyl, butadienyl (including1,2-butadienyl and 1,3-butadienyl).

“Alkynyl” refers to an alkyl group containing at least one carbon-carbontriple bond and having from 2 to 20 carbon atoms (i.e., C₂₋₂₀ alkynyl),2 to 8 carbon atoms (i.e., C₂₋₈ alkynyl), 2 to 6 carbon atoms (i.e.,C₂₋₆ alkynyl) or 2 to 4 carbon atoms (i.e., C₂₋₄ alkynyl). The term“alkynyl” also includes those groups having one triple bond and onedouble bond.

“Alkoxy” refers to the group “alkyl-O—”. Examples of alkoxy groupsinclude, e.g., methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy,tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy and 1,2 dimethylbutoxy.

“Alkoxyalkyl” refers to the group “alkyl-O-alkyl”.

“Amino” refers to the group —NR^(y)R^(z) wherein R^(y) and R^(z) areindependently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may beoptionally substituted, as defined herein.

“Aryl” refers to an aromatic carbocyclic group having a single ring(e.g., monocyclic) or multiple rings (e.g., bicyclic or tricyclic)including fused systems. As used herein, aryl has 6 to 20 ring carbonatoms (i.e., C₆₋₂₀ aryl), 6 to 12 carbon ring atoms (i.e., C₆₋₁₂ aryl),or 6 to 10 carbon ring atoms (i.e., C₆₋₁₀ aryl). Examples of aryl groupsinclude, e.g., phenyl, naphthyl, fluorenyl and anthryl. Aryl, however,does not encompass or overlap in any way with heteroaryl defined below.If one or more aryl groups are fused with a heteroaryl, the resultingring system is heteroaryl. If one or more awl groups are fused with aheterocyclyl, the resulting ring system is heterocyclyl.

“Cycloalkyl” refers to a saturated or partially unsaturated cyclic alkylgroup having a single ring or multiple rings including fused, bridgedand Spiro ring systems. The term “cycloalkyl” includes cycloalkenylgroups (i.e., the cyclic group having at least one double bond) andcarbocyclic fused ring systems having at least one sp³ carbon atom(i.e., at least one non-aromatic ring). As used herein, cycloalkyl hasfrom 3 to 20 ring carbon atoms (i.e., C₃₋₂₀ cycloalkyl), 3 to 12 ringcarbon atoms (i.e., C₃₋₁₂ cycloalkyl), 3 to 10 ring carbon atoms (i.e.,C₃₋₁₀ cycloalkyl), 3 to 8 ring carbon atoms (i.e., C₃₋₈cycloalkyl), or 3to 6 ring carbon atoms (i.e., C₃₋₆ cycloalkyl). Monocyclic groupsinclude, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl and cyclooctyl. Further, the term cycloalkyl is intended toencompass any non-aromatic ring which may be fused to an awl ring,regardless of the attachment to the remainder of the molecule. Stillfurther, cycloalkyl also includes “spirocycloalkyl” when there are twopositions for substitution on the same carbon atom.

“Heteroaryl” refers to an aromatic group having a single ring, multiplerings or multiple fused rings, with one or more ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. As usedherein, heteroaryl includes 1 to 20 ring carbon atoms (i.e., C₁₋₂₀heteroaryl), 3 to 12 ring carbon atoms (i.e., C₃₋₁₂ heteroaryl), or 3 to8 carbon ring atoms (i.e., C₃₋₈ heteroaryl), and 1 to 5 ringheteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2ring heteroatoms, or 1 ring heteroatom independently selected fromnitrogen, oxygen and sulfur. In certain instances, heteroaryl includes5-10 membered ring systems, 5-7 membered ring systems, or 5-6 memberedring systems, each independently having 1 to 4 ring heteroatoms, 1 to 3ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatomindependently selected from nitrogen, oxygen and sulfur. Any aromaticring, having a single or multiple fused rings, containing at least oneheteroatom, is considered a heteroaryl regardless of the attachment tothe remainder of the molecule (i.e., through any one of the fusedrings). Heteroaryl does not encompass or overlap with awl as definedabove.

“Heterocyclyl” refers to a saturated or partially unsaturated cyclicalkyl group, with one or more ring heteroatoms independently selectedfrom nitrogen, oxygen and sulfur. The term “heterocyclyl” includesheterocycloalkenyl groups (i.e., the heterocyclyl group having at leastone double bond), bridged-heterocyclyl groups, fused-heterocyclyl groupsand spiro-heterocyclyl groups. A heterocyclyl may be a single ring ormultiple rings wherein the multiple rings may be fused, bridged orspiro, and may comprise one or more (e.g., 1 to 3) oxo (═O) or N-oxide(—O⁻) moieties. Any non-aromatic ring containing at least one heteroatomis considered a heterocyclyl, regardless of the attachment (i.e., can bebound through a carbon atom or a heteroatom). Further, the termheterocyclyl is intended to encompass any non-aromatic ring containingat least one heteroatom, which ring may be fused to an aryl orheteroaryl ring, regardless of the attachment to the remainder of themolecule. As used herein, heterocyclyl has 2 to 20 ring carbon atoms(i.e., C₂₋₂₀ heterocyclyl), 2 to 12 ring carbon atoms (i.e., C₂₋₁₂heterocyclyl), 2 to 10 ring carbon atoms (i.e., C₂₋₁₀ heterocyclyl), 2to 8 ring carbon atoms (i.e., C₂₋₈ heterocyclyl), 3 to 12 ring carbonatoms (i.e., C₃₋₁₂ heterocyclyl), 3 to 8 ring carbon atoms (i.e., C₃₋₈heterocyclyl), or 3 to 6 ring carbon atoms (i.e., C₃₋₆ heterocyclyl);having 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ringheteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independentlyselected from nitrogen, sulfur or oxygen. The term “heterocyclyl” alsoincludes “spiroheterocyclyl” when there are two positions forsubstitution on the same carbon atom.

“Alkylene” refers to a divalent alkyl group as defined above. As usedherein, alkylene has 1 to 10 carbon atoms (i.e., C₁₋₁₀ alkylene), 1 to 8carbon atoms (i.e., C₁₋₈ alkylene), 1 to 6 carbon atoms (i.e., C₁₋₆alkylene), or 1 to 4 carbon atoms (i.e., C₁₋₄ alkylene).

“Heteroalkylene” refers to an alkylene group in which one or more of thecarbon atoms (and any associated hydrogen atoms) are each independentlyreplaced with the same or different heteroatomic group. The term“heteroalkylene” includes unbranched or branched saturated chain havingcarbon and heteroatoms. By way of example, 1, 2 or 3 carbon atoms may beindependently replaced with the same or different heteroatomic group.Heteroatomic groups include, but are not limited to, —NH—, —O—, —S—,—S(O)—, —S(O)₂—, and the like. As used herein, heteroalkylene includes 1to 8 carbon atoms, or 1 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to2 heteroatoms, or 1 heteroatom.

“Alkenylene” refers to an alkylene group containing at least onecarbon-carbon double bond and having from 2 to 8 carbon atoms (i.e.,C₂₋₈ alkenylene), 2 to 6 carbon atoms (i.e., C₂₋₆ alkenylene), or 2 to 4carbon atoms (i.e., C₂₋₄ alkenylene).

“Heteroalkenylene” refers to a heteroalkylene group containing at leastone carbon-carbon double bond and having from 2 to 8 carbon atoms, 2 to6 carbon atoms, or 2 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2heteroatoms, or 1 heteroatom. The term “heteroalkynyl” also includesthose groups having one triple bond and one double bond.

“Alkynylene” refers to an alkylene group containing at least onecarbon-carbon triple bond and having from 2 to 8 carbon atoms (i.e.,C₂₋₈ alkynylene), 2 to 6 carbon atoms (i.e., C₂₋₆ alkynylene), or 2 to 4carbon atoms (i.e., C₂₋₄ alkynylene). The term “alkynyl” also includesthose groups having one triple bond and one double bond.

“Heteroalkynylene” refers to a heteroalkylene group containing at leastone carbon-carbon triple bond and having from 2 to 8 carbon atoms, 2 to6 carbon atoms, or 2 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2heteroatoms, or 1 heteroatom. The term “heteroalkynyl” also includesthose groups having one triple bond and one double bond.

“Cycloalkylene” refers to a divalent saturated or partially unsaturatedcyclic alkyl group having a single ring or multiple rings includingfused, bridged, and spiro ring systems. The term “cycloalkylene”includes cycloalkenylene groups (i.e. the cyclic group having at leastone double bond). As used herein, cycloalkenylene has from 3 to 10 ringcarbon atoms (i.e., C₃₋₁₀ cycloalkyl), 3 to 8 ring carbon atoms (i.e.,C₃₋₈ cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C₃₋₆ cycloalkyl).

“Heterocycloalkylene” refers to a cycloalkylene group in which one ormore of the carbon atoms (and any associated hydrogen atoms) are eachindependently replaced with the same or different heteroatomic group. Byway of example, 1, 2 or 3 carbon atoms may be independently replacedwith the same or different heteroatomic group. Heteroatomic groupsinclude, but are not limited to, —NH—, —O—, —S—, —S(O)—, —S(O)₂—, andthe like. As used herein, heterocycloalkylene includes 1 to 9 carbonatoms, or 1 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2heteroatoms, or 1 heteroatom.

“Oxo” refers to ═O.

“Halogen” or “halo” includes fluoro, chloro, bromo, and iodo.

The terms “optional” or “optionally” means that the subsequentlydescribed event or circumstance may or may not occur. The term“optionally substituted” refers to any one or more hydrogen atoms on thedesignated atom or group may or may not be replaced by a moiety otherthan hydrogen.

Provided are also are stereoisomers, mixture of stereoisomers,tautomers, hydrates, solvates, isotopically enriched analog, andpharmaceutically acceptable salts of the compounds described herein.

The compounds disclosed herein, or their pharmaceutically acceptablesalts, may include an asymmetric center and may thus give rise toenantiomers, diastereomers, and other stereoisomeric forms that may bedefined, in terms of absolute stereochemistry, as (R)- or (S)- or, as(D)- or (L)- for amino acids. The present disclosure is meant to includeall such possible isomers, as well as their racemic and optically pureforms. Optically active (+) and (−), (R)- and (S)-, or (D)- and(L)-isomers may be prepared using chiral synthons or chiral reagents, orresolved using conventional techniques, for example, chromatography andfractional crystallization. Conventional techniques for thepreparation/isolation of individual enantiomers include chiral synthesisfrom a suitable optically pure precursor or resolution of the racemate(or the racemate of a salt or derivative) using, for example, chiralhigh pressure liquid chromatography (HPLC). When the compounds describedherein contain olefinic double bonds or other centers of geometricasymmetry, and unless specified otherwise, it is intended that thecompounds include both E and Z geometric isomers.

A “stereoisomer” refers to a compound made up of the same atoms bondedby the same bonds but having different three-dimensional structures,which are not interchangeable. The present disclosure contemplatesvarious stereoisomers and mixtures thereof and includes “enantiomers,”which refers to two stereoisomers whose molecules are nonsuperimposeablemirror images of one another and “diastereomers,” which refers tostereoisomers that have at least two asymmetric atoms, but which are notmirror-images of each other. Thus, all stereoisomers (for example,geometric isomers, optical isomers and the like) of the presentcompounds (including those of the salts, solvates and hydrates of thecompounds), such as those which may exist due to asymmetric carbons onvarious substituents, including enantiomeric forms (which may exist evenin the absence of asymmetric carbons), rotameric forms, atropisomers,and diastereomeric forms, are contemplated.

Diasteromeric mixtures can be separated into their individualdiastereomers on the basis of their physical chemical differences bymethods well known to those skilled in the art, such as, for example, bychromatography and/or fractional crystallization. Enantiomers can beseparated by converting the enantiomeric mixture into a diasteromericmixture by reaction with an appropriate optically active compound (e.g.,chiral auxiliary such as a chiral alcohol or Mosher's acid chloride),separating the diastereomers and converting (e.g., hydrolyzing) theindividual diastereomers to the corresponding pure enantiomers. Also,some of the compounds of Formula (1) may be atropisomers and areconsidered as part of this disclosure. Stereoisomers can also beseparated by use of chiral HPLC.

Some of the compounds exist as tautomers. Tautomers are in equilibriumwith one another. For example, amide containing compounds may exist inequilibrium with imidic acid tautomers. Regardless of which tautomer isshown and regardless of the nature of the equilibrium among tautomers,the compounds are understood by one of ordinary skill in the art tocomprise both amide and imidic acid tautomers. Thus, the amidecontaining compounds are understood to include their imidic acidtautomers. Likewise, the imidic acid containing compounds are understoodto include their amide tautomers.

The term “hydrate” refers to the complex formed by the combining of acompound described herein and water.

A “solvate” refers to an association or complex of one or more solventmolecules and a compound of the disclosure. Examples of solvents thatform solvates include, but are not limited to, water, isopropanol,ethanol, methanol, dimethylsulfoxide, ethylacetate, acetic acid andethanolamine.

Any compound or structure given herein, is also intended to representunlabeled forms as well as isotopically labeled forms of the compounds.These forms of compounds may also be referred to as an “isotopicallyenriched analog.” Isotopically labeled compounds have structuresdepicted herein, except that one or more atoms are replaced by an atomhaving a selected atomic mass or mass number.

Examples of isotopes that can be incorporated into the disclosedcompounds include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine, chlorine and iodine, such as ²H, ³H, ¹¹C, ¹³C,¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I,respectively. Various isotopically labeled compounds of the presentdisclosure, for example those into which radioactive isotopes such as³H, ¹³C and ¹⁴C are incorporated. Such isotopically labelled compoundsmay be useful in metabolic studies, reaction kinetic studies, detectionor imaging techniques, such as positron emission tomography (PET) orsingle-photon emission computed tomography (SPECT) including drug orsubstrate tissue distribution assays or in radioactive treatment ofpatients. Such compounds may exhibit increased resistance to metabolismand are thus useful for increasing the half-life of any compound whenadministered to a mammal, particularly a human. Such compounds aresynthesized by means well known in the art, for example by employingstarting materials in which one or more hydrogens have been replaced bydeuterium.

Certain compounds disclosed herein contain one or more ionizable groups(groups from which a proton can be removed (e.g., —COOH) or added (e.g.,amines) or which can be quaternized (e.g., amines)). All possible ionicforms of such molecules and salts thereof are intended to be includedindividually in the disclosure herein. With regard to salts of thecompounds described herein, one of ordinary skill in the art can selectfrom among a wide variety of available counterions those that areappropriate. In specific applications, the selection of a given anion orcation for preparation of a salt may result in increased or decreasedsolubility of that salt.

As used herein, the term “non-biocleavable linking moiety” is intendedto refer to a linking moiety which is not readily hydrolyzed underphysiological conditions. As used herein, the term “biocleavable linkingmoiety” is intended to refer to a linking moiety which is readilyhydrolyzed under physiological conditions. In certain embodiments, atleast one linking moiety is hydrolyzed under intracellular conditions(e.g., low pH).

As used herein, the term “cancer” refers to a class of diseases ofmammals characterized by uncontrolled cellular growth. The term “cancer”is used interchangeably with the terms “tumor,” “solid tumor,”“malignancy,” “hyperproliferation” and “neoplasm.” Cancer includes alltypes of hyperproliferative growth, hyperplasic growth, neoplasticgrowth, cancerous growth or oncogenic processes, metastatic tissues ormalignantly transformed cells, tissues, or organs, irrespective ofhistopathologic type or stage of invasiveness. Illustrative examplesinclude, lung, prostate, head and neck, breast and colorectal cancer,melanomas and gliomas (such as a high grade glioma, includingglioblastoma multiforme (GBM), the most common and deadliest ofmalignant primary brain tumors in adult humans).

The phrase “solid tumor” includes, for example, lung cancer, head andneck cancer, brain cancer, oral cancer, colorectal cancer, breastcancer, prostate cancer, pancreatic cancer, and liver cancer. Othertypes of solid tumors are named for the particular cells that form them,for example, sarcomas formed from connective tissue cells (for example,bone cartilage, fat), carcinomas formed from epithelial tissue cells(for example, breast, colon, pancreas) and lymphomas formed fromlymphatic tissue cells (for example, lymph nodes, spleen, thymus).Treatment of all types of solid tumors regardless of naming conventionis within the scope of this disclosure.

“Chemotherapeutic agent” refers to any substance capable of reducing orpreventing the growth, proliferation, or spread of a cancer cell, apopulation of cancer cells, tumor, or other malignant tissue. The termis intended also to encompass radiotherapy, or any antitumor oranticancer agent.

As used herein, “treatment” or “treating” is an approach for obtaining abeneficial or desired result, such as a clinical result. For purposes ofthis disclosure, beneficial or desired clinical results include, but arenot limited to, alleviation of a symptom and/or diminishment of theextent of a symptom and/or preventing a worsening of a symptomassociated with a disease or condition. In one variation, beneficial ordesired clinical results include, but are not limited to, alleviation ofa symptom and/or diminishment of the extent of a symptom and/orpreventing a worsening of a symptom associated with a cognitivedisorder, a psychotic disorder, a neurotransmitter-mediated disorderand/or a neuronal disorder. Preferably, treatment of a disease orcondition with a compound of the disclosure or a pharmaceuticallyacceptable salt thereof is accompanied by no or fewer side effects thanare associated with currently available therapies for the disease orcondition and/or improves the quality of life of the individual.

The terms “inhibit,” “inhibiting,” and “inhibition” refer to theslowing, halting, or reversing the growth or progression of a disease,infection, condition, or group of cells. The inhibition can be greaterthan about 20%, 40%, 60%, 80%, 90%, 95%, or 99%, for example, comparedto the growth or progression that occurs in the absence of the treatmentor contacting.

As used herein, by “combination therapy” is meant a therapy thatincludes two or more different compounds. Thus, in one aspect, acombination therapy comprising a compound detailed herein and anthercompound is provided. In some variations, the combination therapyoptionally includes one or more pharmaceutically acceptable carriers orexcipients, non-pharmaceutically active compounds, and/or inertsubstances. In various embodiments, treatment with a combination therapymay result in an additive or even synergistic (e.g., greater thanadditive) result compared to administration of a single compound of thedisclosure alone. In some embodiments, a lower amount of each compoundis used as part of a combination therapy compared to the amountgenerally used for individual therapy. Preferably, the same or greatertherapeutic benefit is achieved using a combination therapy than byusing any of the individual compounds alone. In some embodiments, thesame or greater therapeutic benefit is achieved using a smaller amount(e.g., a lower dose or a less frequent dosing schedule) of a compound ina combination therapy than the amount generally used for individualcompound or therapy. Preferably, the use of a small amount of compoundresults in a reduction in the number, severity, frequency, and/orduration of one or more side-effects associated with the compound.

As used herein, the term “effective amount” intends such amount of acompound of the disclosure which in combination with its parameters ofefficacy and toxicity, as well as based on the knowledge of thepracticing specialist should be effective in a given therapeutic form.As is understood in the art, an effective amount may be in one or moredoses, i.e., a single dose or multiple doses may be required to achievethe desired treatment endpoint. An effective amount may be considered inthe context of administering one or more therapeutic agents, and asingle agent may be considered to be given in an effective amount if, inconjunction with one or more other agents, a desirable or beneficialresult may be or is achieved. Suitable doses of any of theco-administered compounds may optionally be lowered due to the combinedaction (e.g., additive or synergistic effects) of the compounds.

As used herein, the term “agonist” refers to a compound, the presence ofwhich results in a biological activity of a protein that is the same asthe biological activity resulting from the presence of a naturallyoccurring ligand for the protein, such as, for example, PARP.

As used herein, the term “partial agonist” refers to a compound thepresence of which results in a biological activity of a protein that isof the same type as that resulting from the presence of a naturallyoccurring ligand for the protein, but of a lower magnitude.

As used herein, the term “antagonist” or “inhibitor” refers to acompound, the presence of which results in a decrease in the magnitudeof a biological activity of a protein. In certain embodiments, thepresence of an antagonist results in complete inhibition of a biologicalactivity of a protein, such as, for example, the enzyme poly(ADP-ribose)polymerase (PARP).

As used herein, the IC₅₀ refers to an amount, concentration or dosage ofa particular test compound that achieves a 50% inhibition of a maximalresponse, such as modulation of PARP, in an assay that measures suchresponse.

As used herein, EC₅₀ refers to a dosage, concentration or amount of aparticular test compound that elicits a dose-dependent response at 50%of maximal expression of a particular response that is induced, provokedor potentiated by the particular test compound.

The term “cancer,” as used herein refers to an abnormal growth of cellswhich tend to proliferate in an uncontrolled way and, in some cases, tometastasize (spread). The types of cancer include, but are not limitedto, solid tumors (such as those of the bladder, bowel, brain, breast,endometrium, heart, kidney, lung, lymphatic tissue (lymphoma), ovary,pancreas or other endocrine organ (thyroid)), prostate, skin (melanoma)or hematological tumors (such as the leukemias).

The term “carrier,” as used herein, refers to relatively nontoxicchemical compounds or agents that facilitate the incorporation of acompound into cells or tissues.

As used herein, “unit dosage form” refers to physically discrete units,suitable as unit dosages, each unit containing a predetermined quantityof active ingredient calculated to produce the desired therapeuticeffect in association with the required pharmaceutical carrier. Unitdosage forms may contain a single or a combination therapy.

As used herein, the term “controlled release” refers to adrug-containing formulation or fraction thereof in which release of thedrug is not immediate, i.e., with a “controlled release” formulation,administration does not result in immediate release of the drug into anabsorption pool. The term encompasses depot formulations designed togradually release the drug compound over an extended period of time.Controlled release formulations can include a wide variety of drugdelivery systems, generally involving mixing the drug compound withcarriers, polymers or other compounds having the desired releasecharacteristics (e.g., pH-dependent or non-pH-dependent solubility,different degrees of water solubility, and the like) and formulating themixture according to the desired route of delivery (e.g., coatedcapsules, implantable reservoirs, injectable solutions containingbiodegradable capsules, and the like).

As used herein, by “pharmaceutically acceptable” or “pharmacologicallyacceptable” is meant a material that is not biologically or otherwiseundesirable, e.g., the material may be incorporated into apharmaceutical composition administered to a patient without causing anysignificant undesirable biological effects or interacting in adeleterious manner with any of the other components of the compositionin which it is contained. Pharmaceutically acceptable carriers orexcipients have preferably met the required standards of toxicologicaland manufacturing testing and/or are included on the Inactive IngredientGuide prepared by the U.S. Food and Drug administration.

“Pharmaceutically acceptable salts” are those salts which retain atleast some of the biological activity of the free (non-salt) compoundand which can be administered as drugs or pharmaceuticals to anindividual. Such salts, for example, include: (1) acid addition salts,formed with inorganic acids such as hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, phosphoric acid, and the like; or formedwith organic acids such as acetic acid, oxalic acid, propionic acid,succinic acid, maleic acid, tartaric acid and the like; (2) salts formedwhen an acidic proton present in the parent compound either is replacedby a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or analuminum ion; or coordinates with an organic base. Acceptable organicbases include ethanolamine, diethanolamine, triethanolamine and thelike. Acceptable inorganic bases include aluminum hydroxide, calciumhydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, andthe like. Further examples of pharmaceutically acceptable salts includethose listed in Berge et al., Pharmaceutical Salts, J. Pharm. Sci. 1977January; 66(1):1-19. Pharmaceutically acceptable salts can be preparedin situ in the manufacturing process, or by separately reacting apurified compound of the disclosure in its free acid or base form with asuitable organic or inorganic base or acid, respectively, and isolatingthe salt thus formed during subsequent purification. It should beunderstood that a reference to a pharmaceutically acceptable saltincludes the solvent addition forms or crystal forms thereof,particularly solvates or polymorphs. Solvates contain eitherstoichiometric or non-stoichiometric amounts of a solvent, and are oftenformed during the process of crystallization. Hydrates are formed whenthe solvent is water, or alcoholates are formed when the solvent isalcohol. Polymorphs include the different crystal packing arrangementsof the same elemental composition of a compound. Polymorphs usually havedifferent X-ray diffraction patterns, infrared spectra, melting points,density, hardness, crystal shape, optical and electrical properties,stability, and solubility. Various factors such as the recrystallizationsolvent, rate of crystallization, and storage temperature may cause asingle crystal form to dominate.

The term “excipient” as used herein means an inert or inactive substancethat may be used in the production of a drug or pharmaceutical, such asa tablet containing a compound of the disclosure as an activeingredient. Various substances may be embraced by the term excipient,including without limitation any substance used as a binder,disintegrant, coating, compression/encapsulation aid, cream or lotion,lubricant, solutions for parenteral administration, materials forchewable tablets, sweetener or flavoring, suspending/gelling agent, orwet granulation agent. Binders include, e.g., carbomers, povidone,xanthan gum, etc.; coatings include, e.g., cellulose acetate phthalate,ethylcellulose, gellan gum, maltodextrin, enteric coatings, etc.;compression/encapsulation aids include, e.g., calcium carbonate,dextrose, fructose dc (directly compressible), honey dc, lactose(anhydrate or monohydrate; optionally in combination with aspartame,cellulose, or microcrystalline cellulose), starch dc, sucrose, etc.;disintegrants include, e.g., croscarmellose sodium, gellan gum, sodiumstarch glycolate, etc.; creams or lotions include, e.g., maltodextrin,carrageenans, etc.; lubricants include, e.g., magnesium stearate,stearic acid, sodium stearyl fumarate, etc.; materials for chewabletablets include, e.g., dextrose, fructose dc, lactose (monohydrate,optionally in combination with aspartame or cellulose), etc.;suspending/gelling agents include, e.g., carrageenan, sodium starchglycolate, xanthan gum, etc.; sweeteners include, e.g., aspartame,dextrose, fructose dc, sorbitol, sucrose dc, etc.; and wet granulationagents include, e.g., calcium carbonate, maltodextrin, microcrystallinecellulose, etc.

Compounds

Provided herein are compounds comprising at least one nuclear payloadand at least one nuclear receptor-targeting epitope. The compoundsdescribed herein are capable of targeting the nucleus of a cell byrecognition and binding of a nuclear receptor-targeting epitope to therespective binding site and delivering the nuclear payload to thenucleus of the cell. The nuclear payload then is capable of binding toone or more target sites within the nucleus and/or disrupting one ormore cellular processes, causing the cell to die. In certainembodiments, the nuclear payload is bonded to the nuclearreceptor-targeting epitope(s) via a linking moiety. In certainembodiments, the linking moiety provides a single or mono-linkage,meaning that the linker is only conjugated to one atom of each of thepayload and the epitope.

The compounds described herein can comprise more than one nuclearreceptor-targeting epitope. The epitopes can be the same or different,such that the compounds are directed to one or more cellular targets, inaddition to the nucleus.

Accordingly, provided is a compound of Formula I, or stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

A-(L-B)_(m)  I

wherein:

A is a nuclear payload;

m is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope; and

each L is independently a covalent bond or a linking moiety.

In certain embodiments, one or more nuclear receptor-targeting epitopesare bonded to a nuclear payload via a single linking moiety.Accordingly, also provided is a compound of Formula II, or stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

A-L-(B)_(m)  II

wherein:

A is a nuclear payload;

m is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope; and

L is a linking moiety.

In certain embodiments, the nuclear receptor-targeting epitope ofFormula I or TT is a nuclear hormone receptor-targeting epitope. Incertain embodiments, the nuclear receptor-targeting epitope of Formula Ior II is a nuclear steroid receptor-targeting epitope.

Also provided is a compound of Formula I or Formula II, or stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

A-(L-B)_(m)  I

A-L-(B)_(m)  II

wherein:

A is a nuclear payload;

m is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope derivedfrom estrogen, estetrol, estriol, estrone, progesterone, enobosarm,bicalutamide, apalutamide, testosterone, dihydrotestosterone,testosterone, 19-nortestosterone, progesterone, andarine, cortisol,prednisone, estradiol, flutamide, nilutamide, enzalutamide, tamoxifen,toremifene, raloxifene, bazedoxifene, ospemifene, megestrol acetate,estramustine, abiraterone, LGD-2941, BMS-564929, ostarine, ulipristalacetate, asoprisnil (J867), mifepristone, telapristone (CDB-4124,Proellex, Progenta), or an analog thereof; and each L is independently acovalent bond or a linking moiety.

Also provided is a compound of Formula I, or stereoisomer, mixture ofstereoisomers, hydrate, solvate, isotopically enriched analog orpharmaceutically acceptable salt thereof:

A-(L-B)_(m)  I

wherein:

A is a nuclear payload which binds to poly(ADP-ribose) polymerase-1(PARP-1) and/or poly(ADP-ribose) polymerase PARP-2);

m is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope derivedfrom estrogen, estetrol, estriol, estrone, progesterone, enobosarm,bicalutamide, apalutamide, testosterone, dihydrotestosterone, estradiol,flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene,bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone,LGD-2941, BMS-564929, ostarine, or an analog thereof; and

each L is independently a covalent bond or a linking moiety.

Also provided is a compound of Formula II, or stereoisomer, mixture ofstereoisomers, hydrate, solvate, isotopically enriched analog orpharmaceutically acceptable salt thereof:

A-L-(B)_(m)  II

wherein:

A is a nuclear payload which binds to poly(ADP-ribose) polymerase-1(PARP-1) and/or poly(ADP-ribose) polymerase PARP-2);

m is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope derivedfrom estrogen, estetrol, estriol, estrone, progesterone, enobosarm,bicalutamide, apalutamide, testosterone, dihydrotestosterone, estradiol,flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene,bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone,LGD-2941, BMS-564929, ostarine, or an analog thereof; and

each L is independently a covalent bond or a linking moiety.

The “linking moiety” of any compounds described herein can bebiocleavable (e.g., acid labile) or non-biocleavable. Linking moietiescan be linear, branched, saturated, unsaturated, all-carbon orheteroatomic. Linking moieties can also contain one or more rings thatare fused, saturated, unsaturated, as well as be all-carbon orheteroatomic. In certain embodiments, the linking moiety is anon-biocleavable linking moiety. In certain embodiments, the linkingmoiety is a biocleavable linking moiety. In certain embodiments, anuclear payload is bonded to one nuclear steroid receptor-targetingepitope via a non-biocleavable linking moiety and one or more nuclearsteroid receptor-targeting epitope(s) via a biocleavable linking moiety.In certain embodiments, the biocleavable linking moiety is anacid-labile linking moiety. In some embodiments, the linking moietycomprises a hydrazone linkage.

It is contemplated that any linking moiety can be used in the compoundsdescribed herein, provided that it does not significantly interfere withor disrupt the desired binding of the nuclear payload or the nuclearreceptor-targeting epitope. In some embodiments, the linking moiety isalkylene, heteroalkylene, alkenylene, heteroalkenylene, alkynylene,heteroalkynylene, arylene, heteroarylene, cycloalkylene orheterocycloalkylene; wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, may optionally comprisean arylene, heteroarylene, cycloalkylene or heterocycloalkylene; andfurther wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, arylene, heteroarylene,cycloalkylene or heterocycloalkylene is independently optionallysubstituted with one to five substituents independently selected fromoxo, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkyl.

In certain embodiments, the linking moiety is of the formula:

—Y¹—(CH₂)_(n′)—Y²—(CH₂)_(m′)—Y³—

wherein:

each of Y¹, Y², and Y³ are independently a —NR¹¹—, —O—, —S(O)₀₋₂—,—NR¹¹C(O)—, —C(O)NR¹¹—, —NR¹¹S(O)₂—, —S(O)₂NR¹¹—, —CR¹²═N—NR¹¹—,—NR¹¹—N═CR¹²—, —C(O)—, arylene, heteroarylene, cycloalkylene orheterocycloalkylene; wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, arylene, heteroarylene,cycloalkylene or heterocycloalkylene is independently optionallysubstituted with one to five substituents independently selected fromoxo, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkyl;

each R¹¹ is independently C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl, heteroaryl,cycloalkyl or heterocyclyl;

each R¹² is independently C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl, heteroaryl,cycloalkyl or heterocyclyl; and

n′ and m′ are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8.

Also provided is a compound of Formula IA, or stereoisomer, mixture ofstereoisomers, hydrate, solvate, isotopically enriched analog orpharmaceutically acceptable salt thereof:

A-(Y¹—(CH₂)_(p)—Y²—(CH₂)_(q)—Y³-B)_(r)  IA

wherein:

A is a nuclear payload which binds to poly(ADP-ribose) polymerase-1(PARP-1) and/or poly(ADP-ribose) polymerase PARP-2);

r is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope derivedfrom estrogen, estetrol, estriol, estrone, progesterone, enobosarm,bicalutamide, apalutamide, testosterone, dihydrotestosterone, estradiol,flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene,bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone,LGD-2941, BMS-564929, ostarine, or an analog thereof; and

each of Y¹, Y², and Y³ are independently a bond, —NR¹¹—, —O—, —S(O)₀₋₂—,—NR¹¹C(O)—, —C(O)NR¹¹—, —NR¹¹S(O)₂—, —S(O)₂NR¹¹—, —CR¹²═N—NR¹¹—,—NR¹¹—N═CR¹²—, —C(O)—, arylene, heteroarylene, cycloalkylene, orheterocycloalkylene; wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, arylene, heteroarylene,cycloalkylene or heterocycloalkylene is independently optionallysubstituted with one to five substituents independently selected fromoxo, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkyl;

each R¹¹ is independently C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl, heteroaryl,cycloalkyl or heterocyclyl;

each R¹² is independently C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl, heteroaryl,cycloalkyl or heterocyclyl; and

p and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8.

Also provided is a compound of Formula IIA, or stereoisomer, mixture ofstereoisomers, hydrate, solvate, isotopically enriched analog orpharmaceutically acceptable salt thereof:

A-Y¹—(CH₂)_(p)—Y²—(CH₂)_(q)—Y³-(B)_(r)  IIA

wherein:

A is a nuclear payload which binds to poly(ADP-ribose) polymerase-1(PARP-1) and/or poly(ADP-ribose) polymerase PARP-2);

r is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope derivedfrom estrogen, estetrol, estriol, estrone, progesterone, enobosarm,bicalutamide, apalutamide, testosterone, dihydrotestosterone, estradiol,flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene,bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone,LGD-2941, BMS-564929, ostarine, or an analog thereof; and

each of Y¹, Y², and Y³ are independently a bond, —NR¹¹—, —O—, —S(O)₀₋₂—,—NR¹¹C(O)—, —C(O)NR¹¹, —NR¹¹S(O)₂—, —S(O)₂NR¹¹—, —CR¹²═N—NR¹¹—,—NR¹¹—N═CR¹²—, —C(O)-arylene, heteroarylene, cycloalkylene orheterocycloalkylene; wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, arylene, heteroarylene,cycloalkylene or heterocycloalkylene is independently optionallysubstituted with one to five substituents independently selected fromoxo, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkyl;

each R¹¹ is independently C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl, heteroaryl,cycloalkyl or heterocyclyl; each R¹² is independently C₁₋₄ alkyl, C₁₋₄haloalkyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; and

p and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8.

In certain embodiments, the linking moiety is not a bond. In certainembodiments, each R¹¹ is independently hydrogen, C₁₋₄ alkyl, C₁₋₄haloalkyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; and each R¹² isindependently hydrogen, C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl, heteroaryl,cycloalkyl or heterocyclyl.

In certain embodiments, the linking moiety is of the formula:

—Y¹—(CH₂)_(n′)—Y²—(CH₂)_(m′)—Y³—

wherein:

each of Y¹, Y², and Y³ are independently a —NR¹¹—, —O—, —S(O)₀₋₂—,—NR¹¹C(O)—, —C(O)NR¹¹—, —NR¹¹S(O)₂—, —S(O)₂NR¹¹—, —CR¹²═N—NR¹¹—,—NR¹¹—N═CR¹²—, —C(O)—, arylene, heteroarylene, cycloalkylene orheterocycloalkylene; wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, arylene, heteroarylene,cycloalkylene or heterocycloalkylene is independently optionallysubstituted with one to five substituents independently selected fromoxo, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkyl;

each R¹¹ is independently hydrogen, C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl,heteroaryl, cycloalkyl or heterocyclyl;

each R¹² is independently hydrogen, C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl,heteroaryl, cycloalkyl or heterocyclyl; and

n′ and m′ are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8.

Also provided is a compound of Formula I or II, or a stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

A-(L-B)_(m)  I

A-L-(B)_(m)  II

wherein:

A is a nuclear payload;

m is 1, 2 or 3;

each B is independently a nuclear receptor-targeting epitope; and

each L is independently a covalent bond or a linking moiety.

Also provided is a compound of Formula I or II, or a stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

A-(L-B)_(m)  I

A-L-(B)_(m)  II

wherein:

A is a nuclear payload;

m is 1, 2 or 3;

each B is independently a nuclear steroid receptor-targeting epitope;and

each L is independently a covalent bond or a linking moiety.

In certain embodiments of any Formula or subformula disclosed herein(e.g., Formula I or II), when m is 1, then the nuclearreceptor-targeting epitope is not a peptide, protein, nanoparticle orantibody. In certain embodiments of any Formula or subformula disclosedherein (e.g., Formula I or II), when m is 1, and B is an androgenreceptor-targeting epitope, then A is not doxorubicin, or an analogthereof. In certain embodiments of any Formula or subformula disclosedherein (e.g., Formula I or II), when m is 1, and B is an androgenreceptor-targeting epitope, then A is not a hydroxamic acid which bindshistone deacetylase (HDAC). In certain embodiments of any Formula orsubformula disclosed herein (e.g., Formula I or II), when in is 1, and Bis an estrogen receptor-targeting epitope, then A is not doxorubicin, oran analog thereof. In certain embodiments of any Formula or subformuladisclosed herein (e.g., Formula I or II), when m is 1, and B is anestrogen receptor-targeting epitope, then A is not a hydroxamic acidwhich binds histone deacetylase (HDAC).

Also provided is compound of Formula III, IV, V or VI, or stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

wherein:

each R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶,—NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵,wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R¹, R², R³ and R⁴ are independently optionally substitutedwith one or more (e.g., 1 to 5, or 1 to 3) R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more (e.g., 1 to 5, or 1 to 3) haloor C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino asvalency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₅₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino;

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino; and

R⁹ is hydrogen or R²;

provided that at least one R¹, R², R³ and R⁴ is -L-(B)_(m).

Also provided is compound of Formula IIIA, IV, VA or VI, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶,—NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵,wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R¹, R², R³ and R⁴ are independently optionally substitutedwith one or more (e.g., 1 to 5, or 1 to 3) R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more (e.g., 1 to 5, or 1 to 3) haloor C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino asvalency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

Also provided is compound of Formula IIIB, IVA, VB or VIA, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

R² is -L-(B)_(m);

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

Also provided is compound of Formula III′, IV′, V′ or VI′, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶,—NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵,wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R¹, R², R³ and R⁴ are independently optionally substitutedwith one or more (e.g., 1 to 5, or 1 to 3) R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear steroid receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl,

-   -   alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl        of R¹⁰ are independently optionally substituted with one or more        (e.g., 1 to 5, or 1 to 3) halo or C₁₋₁₂ alkyl optionally        substituted by oxo, halo, hydroxyl or amino as valency permits;        and

each R⁵ and R⁶ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁵ and R⁶ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

Also provided is compound of Formula IIIA′, IVA′, VA′ or VIA′, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

R² is -L-(B)_(m);

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear steroid receptor-targeting epitope.

Nuclear Payloads

The nuclear payloads as used herein are generally capable of binding toany site which is involved in a cellular process important for thedevelopment of cancer, or cellular replication. In certain embodiments,the nuclear payload binds to the target site within the nucleus anddisrupts one or more cellular processes, causing the cell to die. Targetsites within the nucleus include, but are not limited to, a sub-nuclearcompartment (e.g., promyelocytic leukemia nuclear body (PML NB),nucleolus), a protein-protein interaction within the nucleus (e.g.,hypoxia-inducible factor 1α (HIF-1α), FKBP25) or modifications of thechromatin structure. In certain embodiments, the nuclear payload targetsa protein involved in the DNA damage repair process, such as, but notlimited to, poly(ADP-ribose) polymerase (PARP), DNA-dependent proteinkinase (DNA-PK), myelin transcription factor 1 (MYT1), p53,melanocyte-stimulating hormone (MSH), mutL homolog (MLH), ERCC1,apurinic/apyrimidinic endonuclease 1 (APE1), topoisomerase I (Topo I),topoisomerase II (Topo II), Wee1, checkpoint kinase1 (Chk1), checkpointkinase2 (Chk2), ataxia telangiectasia (ATR), or ataxia-telangiectasiamutated (ATM).

In certain embodiments, the nuclear payload comprises olaparib(AZD-2281), Olaparib TOPARP-A, rucaparib (AG014699, PF-01367338),niraparib, talazoparib (BMN-673), veliparib (ABT-888), CEP 9722, E7016,BGB-290, 3-aminobenzamide, methoxyamine, CC-115, MSC2490484A, AZD6738,VX-970, AZD0156, GDC-0575, MK-8776, LY2606368, AZD1775, belotecan,CRLX101, irinotecan, LMP 400, LMP 776, NKTR-102, topotecan, doxorubicin,epirubicin, etoposide, idarubicin, mitoxantrone, teniposide, or ananalog thereof. In certain embodiments, the nuclear payload comprisesAZD-1775 (MK-1775, Adavosertib), SCH900776 (MK-8776), AZD0156, M6620(VX-970, VE-822, Berzosertib), AZD6738, or CC-115, or an analog thereof.In certain embodiments, the nuclear payload comprises a combination ofCC-115 with an additional nuclear payload. In certain embodiments, thenuclear payload comprises CC-115 and the compound comprises enzalutamideor an analog thereof.

The analogs are derived from the known nuclear payloads named herein andare modified to be conjugated to at least one nuclear receptor-targetingepitope, optionally via a linking moiety. The analogs, even aftermodification to arrive at the compounds described herein, maintainbiological activity, which is comparable to that observed in theoriginal, unmodified nuclear payload. In certain embodiments, theanalogs exhibit a binding activity or inhibition which is at least about98%, about 95%, about 90%, about 85%, about 80%, about 75%, about 70%,about 65%, about 60%, about 55%, or about 50% of that observed in theoriginal, unmodified nuclear payload.

In certain embodiments, the terms “modified” and “derived from” as usedin reference to a nuclear payload, means that at most, one non-hydrogenatom of the original, unmodified nuclear payload (i.e., a known nuclearpayload) is replaced by a covalent bond linking at least one nuclearreceptor-targeting epitope(s), optionally via a linking moiety. Incertain embodiments, the terms “modified” and “derived from” as used inreference to a nuclear payload, means that at most, only one hydrogenatom of the original, unmodified nuclear payload (i.e., a known nuclearpayload) is replaced by a covalent bond linking at least one nuclearreceptor-targeting epitope(s), optionally via a linking moiety. Incertain embodiments, one hydrogen atom bound to a heteroatom (e.g., N,O, or S) of the original, unmodified nuclear payload (i.e., a knownnuclear payload) is replaced by a covalent bond linking at least onenuclear receptor-targeting epitope(s), optionally via a linking moiety.

In certain embodiments, the nuclear payload binds to an epigenetictarget, such as histone deacetylase (HDAC) (e.g., vorinostat, romidepsin(Istodax), chidamide, panobinostat (Farydak), belinostat (PXD101),panobinostat (LBH589), valproic acid (as Mg valproate), mocctinostat(MGCD0103), abexinostat (PCI-24781), entinostat (MS-275), SB939,resminostat (4SC-201), givinostat (ITF2357), quisinostat (JNJ-26481585),HBI-8000, kevetrin, CUDC-101, AR-42, CHR-2845, CHR-3996, 4SC-202,CG200745, ACY-1215, ME-344, sulforaphane, etc., or an analog thereof),enhancer of zeste homolog 2 (EZH2) (e.g., tazemetostat, MAK638,CPI-1205), DS-3201b, etc., or an analog thereof), histone acetyltransferase (HAT) (e.g., anacardic acid, MG149, C646, etc., or an analogthereof), methyltransferase (e.g., S-adenosyl methionine, etc., or ananalog thereof), a bromodomain (e.g., JQ1, I-BET 151 (GSK1210151A),I-BET 762 (GSK525762), OTX-015, TEN-010, CPI-203, CPI-0610, olinone,LY294002, or an analog thereof), and the like.

Any known nuclear payload which targets proteins one or more cellularprocesses can be used as the nuclear payload of the compounds describedherein. Small molecule nuclear payloads (i.e., molecular weight of lessthan about 1,000 g/mol) are contemplated to be especially useful in thecompounds described herein (e.g., tripazamine, chetomin, rapamycin, PARPinhibitors, etc.).

In certain embodiments, the compound comprises at least one nuclearpayload which binds to poly(ADP-ribose) polymerase (PARP) and arereferred to herein as “PARP inhibitors.” PARP inhibitors are cytotoxicagents that prevent such DNA repair leading to the death of cells andtumor growth inhibition. In certain embodiments, the PARP is human PARP,and comprises PARP-1 and/or PARP-2, or a variant thereof. In certainembodiments, the nuclear payload is capable of blocking the enzymaticactivity of PARP and/or localizing PARP proteins to sites of DNA damage(i.e., “PARP trapping”). Accordingly, in certain embodiments, thenuclear payload binds to PARP and induces an allosteric conformationalchange in the enzyme.

In certain embodiments, the nuclear payload binds to the PARP-1catalytic domain. In certain embodiments, the nuclear payload binds tothe PARP-2 catalytic domain. In certain embodiments, the nuclear payloadbinds to a conserved HYE motif. In certain embodiments, the nuclearpayload binds to the nicotinamide-binding pocket in the PARP protein.

In one embodiment, the nuclear payload is an analog of a known PARPinhibitor. Exemplary PARP inhibitors which can be used as nuclearpayloads in the compounds described herein include, but are not limitedto olaparib (AZD-2281), olaparib TOPARP-A, rucaparib (AG014699,PF-01367338), niraparib, talazoparib (BMN-673), veliparib (ABT-888), CEP9722, E7016, BGB-290, and 3-aminobenzamide, or an analog thereof.

The PARP inhibitor analogs are derived from PARP inhibitors and aremodified to be conjugated to at least one nuclear steroidreceptor-targeting epitope, optionally via a linking moiety. The PARPinhibitor analogs, even after modification to arrive at the compoundsdescribed herein, maintain biological activity which is comparable tothat observed in the original, unmodified PARP inhibitor. In certainembodiments, the PARP inhibitor analogs maintain the ability to inhibitPARP. In certain embodiments, the PARP inhibitor analogs exhibit abinding activity which is at least about 98%, about 95%, about 90%,about 85%, about 80%, about 75%, about 70%, about 65%, about 60%, about55%, or about 50% of that observed in the original, unmodified PARPinhibitor. In certain embodiments, the compound as described herein isbinds to a poly(ADP-ribose) polymerase (PARP) (e.g., PARP-1 and/orPARP-2) with an IC₅₀ of less than about 500 nM, or less than about 400nM, or less than about 350 nM, or less than about 300 nM, or less thanabout 200 nM, or less than about 100 nM, or less than about 50 nM.

In certain embodiments, the nuclear payload (e.g., PARP inhibitoranalog) comprises one or more moieties capable of having a bindinginteraction with G863, Y907, S904, A898, K903, E988, Y896, and/or Y889of PARP-1. In certain embodiments, the nuclear payload (e.g., PARPinhibitor analog) comprises one or more moieties capable of having abinding interaction with Y889, Y896, H862, G863, S904, Y907, K903, E988,and/or M890 of PARP-1. In certain embodiments, the nuclear payload(e.g., PARP inhibitor analog) comprises one or more moieties capable ofhaving a binding interaction with Y896, Q763, G863, S904, Y907, K903,and/or E988 of PARP-1. In certain embodiments, the nuclear payload(e.g., PARP inhibitor analog) comprises one or more positively chargedmoieties (e.g., amino group) which interact with the side chains ofQ763, D766, and/or Y896 of PARP-1. In certain embodiments, the nuclearpayload (e.g., PARP inhibitor analog) comprises one or more moietiescapable of having a binding interaction with E322, D326, I425, S417,H415, E545, and/or Y449 of PARP-2.

In certain embodiments, the nuclear payload comprises rucaparib(AG014699, PF-01367338), or an analog thereof (i.e.,rucaparib-containing analogs). Accordingly, provided is a compound ofFormula III, or stereoisomer, mixture of stereoisomers, hydrate,solvate, isotopically enriched analog or pharmaceutically acceptablesalt thereof:

wherein:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR'S(═O)₁₋₂R⁸, —NR'S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino as valency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, m is 1. In certain embodiments, m is 2. Incertain embodiments, only one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, provided is a compound of Formula IIIA, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁶ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino as valency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, provided is a compound of Formula IIIB, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

R² is -L-(B)_(m);

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

In certain embodiments of Formula III, IIIA and IIIB, the nuclearreceptor-targeting epitope is a nuclear steroid receptor-targetingepitope. In certain embodiments of Formula III, IIIA and IIIB, m is 1.In certain embodiments of Formula III, IIIA and IIIB, m is 2. In certainembodiments of Formula III and IIIA, only one of R¹, R², R³ and R⁴ is-L-(B)_(m).

In certain embodiments, the nuclear payload is derived from talazoparib(BMN-673), or an analog thereof (i.e., talazoparib-containing analogs).Accordingly, provided is a compound of Formula IV, or stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

wherein:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino as valency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, provided is a compound of Formula IVA, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

In certain embodiments, provided is a compound of Formula IVB, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

R²¹ and R²³ are each independently selected from hydrogen, halo,hydroxyl, C₁₋₁₂ alkyl, C₃₋₁₀ cycloalkyl, C₁₋₁₂ alkoxy, C₁₋₁₂alkoxyalkyl; wherein each alkyl, cycloalkyl, alkoxy, alkoxyalkyl areindependently optionally substituted with at least one substituentselected from cyano, halo, hydroxyl, nitro, C₁₋₁₂ alkyl, and C₃₋₁₀cycloalkyl, wherein R²³ is not hydroxyl;

R²² and R²⁴ are each independently hydrogen, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl,C₂₋₁₂alkynyl, C₁₋₁₂ alkoxy,

-   -   C(═O)R⁵⁰, —C(═O)OR⁵⁰, —C(═O)N(R⁵⁰)₂, —S(═O)₀₋₂R⁵⁰,        —S(═O)₁₋₂N(R⁵⁰)₂, —NR⁵⁰S(═O)₁₋₂R⁵⁰, C₃₋₁₀ cycloalkyl, aryl,        heterocyclyl, or heteroaryl, wherein each C₁₋₁₂ alkyl, C₂₋₁₂        alkenyl, C₂₋₁₂ alkynyl, C₁₋₁₂ alkoxy, C₃₋₁₀ cycloalkyl, aryl,        heterocyclyl, or heteroaryl may be independently optionally        substituted with 1, 2, or 3 R²⁹;

R²⁰ and R²⁵ are each independently selected from the group consisting ofhydrogen, C₁₋₁₂ alkyl, C₃₋₁₀ cycloalkyl, C₁₋₁₂ alkoxyalkyl, C₁₋₁₂haloalkyl, C₁₋₁₂ alkyl-OH and C₁₋₁₂ alkyl-NR⁵¹R⁵²;

R²⁶, R²⁷, and R²⁸ are each independently selected from the groupconsisting of hydrogen, halo, cyano, nitro, amino, hydroxyl, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₁₂ alkoxy, C₃₋₁₀ cycloalkyl,C(═O)-alkyl, —C(═O)-alkoxy, haloalkoxy, haloalkyl, heteroalkyl; whereineach alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R¹⁰ are independently optionally substituted with one ormore halo, hydroxyl, or C₁₋₁₂ alkyl optionally substituted by oxo, halo,hydroxyl or amino as valency permits;

each R²⁹ is selected from hydroxyl, halo, cyano, nitro, —OR⁵¹, —SR⁵¹,—SF₅, —NR⁵¹R⁵², C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀cycloalkyl, heterocyclyl, aryl, heteroaryl, —C(═O)R⁵¹, —C(═O)OR⁵¹,—OC(═O)OR⁵¹, —OC(═O)R⁵¹, —C(═O)NR⁵¹R⁵², —OC(═O)NR⁵¹R⁵²,—NR⁵¹C(═O)NR⁵¹R⁵², —S(═O)₁₋₂R⁵¹, —S(═O)₁₋₂NR⁵¹R⁵², —NR⁵¹S(═O)₁₋₂R⁵²,—NR⁵¹S(═O)₁₋₂NR⁵¹R⁵², —NR⁵¹C(═O)R⁵², —NR⁵¹C(═O)OR⁵² or —C═NOR⁵¹, whereineach alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R²⁹ are independently optionally substituted with one ormore halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxylor amino as valency permits;

each R⁵⁰ is independently hydrogen, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂alkynyl, C₃₋₁₀ cycloalkyl, aryl, heterocyclyl, or heteroaryl, whereineach alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R⁵⁰ are independently optionally substituted with one ormore halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxylor amino as valency permits; and

each R⁵¹ and R⁵² is independently hydrogen, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl,C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, aryl, or heteroaryl,wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R⁵¹ and R⁵² are optionally substituted with oxo, halo,hydroxyl or amino as valency permits; or R⁵¹ and R⁵² are taken togetherwith the atoms to which they are attached to form heterocyclyloptionally substituted by halo or C₁₋₁₂ alkyl optionally substituted byoxo, halo, hydroxyl or amino; provided at least one R²² or R²⁴ comprisesa -L-(B)_(m) group bonded thereto, wherein each L is independently acovalent bond or a linking moiety, each B is independently a nuclearreceptor-targeting epitope, and m is 1, 2 or 3.

In the compounds of Formula VIB, at least one R²² or R²⁴ comprises asubstituent having a -L-(B)_(m) group bonded thereto. As such, it shouldbe understood that the -L-(B)_(m) group is not bound directly to thetricyclic core.

In certain embodiments of Formula IV, IVA and IVB the nuclearreceptor-targeting epitope is a nuclear steroid receptor-targetingepitope. In certain embodiments of Formula IV, IVA and IVB, m is 1. Incertain embodiments of Formula IV, IVA and IVB, m is 2. In certainembodiments of Formula IV, only one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, the nuclear payload is derived from olaparib(AZD-2281), or an analog thereof (i.e., olaparib-containing analogs). Incertain embodiments, the compound is of Formula V, or stereoisomer,mixture of stereoisomers, hydrate, solvate, isotopically enriched analogor pharmaceutically acceptable salt thereof:

wherein:

each R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino as valency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₅₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino;

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

R⁹ is hydrogen or R²; and

provided that at least one R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, provided is a compound of Formula VA, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more R¹⁰ as valency permits;

m is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino as valency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, provided is a compound of Formula VB, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

R² is -L-(B)_(m);

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

In certain embodiments, provided is a compound of Formula VC, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

A and B together represent an optionally substituted, fused aromaticring:

R³⁰ and R³¹ are independently hydrogen or C₁₋₁₂ alkyl, or when X is—CR³³R³⁴, R³⁰, R³¹, R³³ and R³⁴ together with the carbon atoms to whichthey are attached, may form an optionally substituted fused aromaticring;

R³² is hydrogen or halo;

X is —NR³³ or —CR³³R³⁴; where if X is —NR³³ then t is 1 or 2; and if Xis —CR³³R³⁴ then t is 1; R³³ is hydrogen, optionally substituted C₁₋₁₂alkyl, aryl, heterocyclyl, —C(═O)R⁵⁰, —C(═O)OR⁵⁰, —C(═O)N(R⁵⁰)₂,—S(═O)₀₋₂R⁵⁰, —S(═O)₁₋₂N(R⁵⁰)₂, —NR⁵⁰S(═O)₁₋₂R⁵⁰;

R³⁴ is hydrogen, hydroxyl, or amino;

or R³³ and R³⁴ may together form a C₃₋₁₀ cycloalkyl or heterocyclylgroup; and

each R⁵⁰ is independently hydrogen, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂alkynyl, C₃₋₁₀ cycloalkyl, aryl, heterocyclyl, or heteroaryl, whereineach alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R⁵⁰ are independently optionally substituted with one ormore halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxylor amino as valency permits;

provided at least one R³⁰, R³¹, R³³ or R³⁴ group comprises a -L-(B)_(m)group bonded thereto, wherein each L is independently a covalent bond ora linking moiety, each B is independently a nuclear receptor-targetingepitope, and m is 1, 2 or 3.

In certain embodiments of Formula V, VA, VB, and VC, the nuclearreceptor-targeting epitope is a nuclear steroid receptor-targetingepitope. In certain embodiments of Formula V, VA, VB, and VC, m is 1. Incertain embodiments of Formula V, VA, VB, and VC, m is 2. In certainembodiments of Formula V and VA, only one of R¹, R², R³ and R⁴ is-L-(B)_(m).

In certain embodiments, the nuclear payload is derived from veliparib(ABT-888), or an analog thereof (i.e., veliparib-containing analogs).Also provided is a compound of Formula VI, or stereoisomer, mixture ofstereoisomers, hydrate, solvate, isotopically enriched analog orpharmaceutically acceptable salt thereof:

wherein:

each of R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶, —NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more R¹⁰ as valency permits;

in is 1, 2, or 3;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR′R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino as valency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, provided is a compound of Formula VIA, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

R² is -L-(B)_(m);

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

In certain embodiments of Formula VI and VIA, the nuclearreceptor-targeting epitope is a nuclear steroid receptor-targetingepitope. In certain embodiments of Formula VI and VIA, m is 1. Incertain embodiments of Formula VI and VIA, m is 2. In certainembodiments of Formula V, only one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments, the nuclear payload comprises CC-115 or ananalog thereof (CC-115-containing analogs). Also provided is a compoundof Formula VII or stereoisomer, mixture of stereoisomers, hydrate,solvate, isotopically enriched analog or pharmaceutically acceptablesalt thereof:

each of R¹, R² and R³ are independently -L-(B)_(m), hydrogen, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl,aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —OC(═O)R⁵, —C(═O)NR⁵R⁶,—NR⁵C(═O)R⁶, —S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵,wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl andheteroaryl of R¹, R² and R³ are independently optionally substitutedwith one or more R¹⁰ as valency permits;

each L is independently a covalent bond or a linking moiety;

each B is independently a nuclear receptor-targeting epitope;

each R¹⁰ is independently halo, cyano, nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁷, —C(═O)OR⁷, —OC(═O)OR⁷,—OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸, —NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷,—S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸, —NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸,—NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein each alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl and heteroaryl of R¹⁰ are independentlyoptionally substituted with one or more halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino as valency permits; and

each R⁵ and R⁶ is independently hydrogen, deuterium, C₁₋₁₂ alkyl orC₃₋₁₂ cycloalkyl, optionally substituted with oxo, halo, hydroxyl oramino as valency permits; or R⁵ and R⁶ are taken together with the atomsto which they are attached to form heterocyclyl optionally substitutedby halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl oramino; and

each R⁷ and R⁸ is independently hydrogen, deuterium or C₁₋₁₂ alkyloptionally substituted with oxo, halo, hydroxyl or amino as valencypermits; or R⁷ and R⁸ are taken together with the atoms to which theyare attached to form heterocyclyl optionally substituted by halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino;

provided that at least one of R¹, R² and R³ is -L-(B)_(m).

In certain embodiments, provided is a compound of Formula VIIA, orstereoisomer, mixture of stereoisomers, hydrate, solvate, isotopicallyenriched analog or pharmaceutically acceptable salt thereof:

wherein:

R² is -L-(B)_(m);

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

In certain embodiments of Formula VII and VIIA, the nuclearreceptor-targeting epitope is a nuclear steroid receptor-targetingepitope. In certain embodiments of Formula VII and VIIA, m is 1. Incertain embodiments of Formula VII and VILA, m is 2. In certainembodiments of Formula VII, only one of R¹, R², R³ and R⁴ is -L-(B)_(m).

In certain embodiments of a Formula disclosed herein, each R⁵ and R⁶ isindependently hydrogen, deuterium, or C₁₋₁₂ alkyl optionally substitutedwith oxo, halo, hydroxyl or amino as valency permits; or R⁵ and R⁶ aretaken together with the atoms to which they are attached to formheterocyclyl optionally substituted by halo or C₁₋₁₂ alkyl optionallysubstituted by oxo, halo, hydroxyl or amino.

In certain embodiments, the nuclear payload binds DNA-dependent proteinkinase (DNA-PK). In certain embodiments, the nuclear payload is aninhibitor of DNA-dependent protein kinase (DNA-PK). In certainembodiments, the nuclear payload is derived from AZD-1775 (MK-1775,Adavosertib), SCH900776 (MK-8776), AZD0156, M6620 (VX-970, VE-822,Berzosertib), AZD6738, or CC-115, or an analog thereof. In certainembodiments, provided is a compound of formula:

wherein one hydrogen atom is replaced by -L-(B)_(m); wherein

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

In certain embodiments, the hydrogen atom replaced by -L-(B)_(m) is on aheteroatom. In certain embodiments, the hydrogen atom replaced by-L-(B)_(m) is on a nitrogen. In certain embodiments, the hydrogen atomreplaced by -L-(B)_(m) is on an oxygen. In certain embodiments, thehydrogen atom replaced by -L-(B)_(m) is on a carbon. In certainembodiments, the nuclear receptor-targeting epitope is a nuclear steroidreceptor-targeting epitope. In certain embodiments, m is 1. In certainembodiments, m is 2.

In certain embodiments, provided is a compound of Formula:

wherein:

R² is -L-(B)_(m);

m is 1, 2, or 3;

L is a covalent bond or a linking moiety; and

each B is independently a nuclear receptor-targeting epitope.

In certain embodiments of any compound, Formula or embodiment disclosedherein, the nuclear receptor-targeting epitope is a nuclear steroidreceptor-targeting epitope. In certain embodiments, m is 1. In certainembodiments, m is 2.

Nuclear Receptor-Targeting Epitopes

As used herein, “nuclear receptor-targeting epitope” refers to theportion of the compound described herein (e.g., the “B” moiety ofFormula I or Formula II) which portion is derived from a nucleartargeting agent as disclosed herein and interacts with a ligand-bindingdomain of the target nuclear receptor, i.e., the portion of the compoundwhich drives a ligand-binding interaction. The nuclearreceptor-targeting epitope serves to associate the compound with atarget nuclear receptor, e.g. a nuclear steroid receptor, facilitate thelocalization of compound to nuclear steroid receptor-expressing cells,and translocate the nuclear payload from the cytosol to nucleus,allowing the compound to accumulate in the nucleus. The level ofaccumulation can be controlled by selecting the appropriate nuclearreceptor-targeting epitope. For example, the compounds described hereincan accumulate in the nucleus to varying degrees, high in the case of afull agonist (e.g., dihydrotestosterone (DHT)), moderate in the case ofa partial agonist (e.g., bicalutamide), and low, in the case ofantagonists (e.g., enzalutamide), through nuclear translocation of thenuclear steroid receptor which happens, following epitope binding to thereceptor.

The steroid receptor target can be any steroid receptor, including, butnot limited to, those which are over-expressed on cancer cells. Incertain embodiments, at least one nuclear steroid receptor-targetingepitope is capable of binding to a ligand binding domain of a nuclearsteroid receptor, such as a ligand binding domain on an estrogenreceptor, glucocorticoid receptor, progesterone receptor or androgenreceptor.

Exemplary nuclear steroid receptor-targeting epitopes include thosederived from an androgen receptor agonist, an androgen receptorantagonist, a selective androgen-receptor modulator (SARM), an estrogenreceptor agonist, an estrogen receptor antagonist, a selective estrogenreceptor modulator (SERM), a glucocorticoid receptor antagonist, aglucocorticoid receptor agonist, a selective glucocorticoid receptormodulator (SGRM), a progesterone receptor antagonist, a progesteronereceptor agonist, a selective progesterone receptor modulator (SPRM), ora combination thereof. The nuclear steroid receptor-targeting epitopesare typically capable of binding to a nuclear steroid receptor with anIC₅₀ of less than about 500 nM, or less than about 400 nM, or less thanabout 300 nM, or less than about 200 nM, or less than about 100 nM, orwith an EC₅₀ of less than about 1 μM, or less than about 900 nM, or lessthan about 800 nM, or less than about 700 nM, or less than about 600 nM,or less than about 500 nM, or less than about 400 nM, or less than about3400 nM, or less than about 200 nM, or less than about 100 nM.

In certain embodiments, the nuclear steroid receptor-targeting epitopeis an agonist at the androgen receptor. In certain embodiments, thenuclear steroid receptor-targeting epitope is an antagonist at theandrogen receptor.

In certain embodiments, the nuclear steroid receptor-targeting epitopeis steroidal (e.g., dihydrotestosterone). In certain embodiments, thenuclear steroid receptor-targeting epitope is non-steroidal (e.g.,enzalutamide, apalutamide and bicalutamide).

The analogs are derived from the known nuclear steroidreceptor-targeting epitope described herein and are modified to beconjugated to at least one nuclear steroid payload, optionally via alinking moiety. The analogs, even after modification to arrive at thecompounds described herein, maintain biological activity, which iscomparable to that observed in the original, unmodified nuclear steroidreceptor-targeting epitope. In certain embodiments, the analogs exhibita binding activity or inhibition which is at least about 98%, about 95%,about 90%, about 85%, about 80%, about 75%, about 70%, about 65%, about60%, about 55%, or about 50% of that observed in the original,unmodified nuclear steroid receptor-targeting epitope.

In certain embodiments, the analogs are derived from a known nuclearreceptor-targeting epitope, such as a known nuclear steroidreceptor-targeting epitope. In certain embodiments, the term “derivedfrom” as used in reference to a nuclear receptor-targeting epitope,means that at most, one non-hydrogen atom of an original, unmodifiednuclear receptor-targeting compound (i.e., a known nuclear steroidreceptor-targeting compound) is replaced by a covalent bond to thenuclear payload, optionally via a linking moiety. Exemplary non-hydrogenatoms include, but are not limited to, —CH₃, —OH, ═O, and —NHS. Incertain embodiments, the term “derived from” as used in reference to anuclear receptor-targeting epitope, means that at most, one non-hydrogenatom of an original, unmodified nuclear receptor-targeting compound(i.e., a known nuclear steroid receptor-targeting compound) is replacedby a covalent bond to the nuclear payload, optionally via a linkingmoiety. In certain embodiments, one hydrogen atom bound to a heteroatom(e.g., N, O, or S) of the original, unmodified nuclearreceptor-targeting compound (i.e., a known nuclear steroidreceptor-targeting compound) is replaced by a covalent bond to thenuclear payload, optionally via a linking moiety.

In certain embodiments, the nuclear steroid receptor-targeting epitopeis an androgen receptor-targeting epitope. As used herein, the term“androgen receptor-targeting epitope” is intended to refer to theportion of the compound which binds to an androgen receptor agonist orandrogen receptor antagonist (including partial androgen receptoragonists or partial androgen receptor antagonists) and which is capableof shuttling a compound from the cytoplasm into the nucleus of a cell.The “androgen receptor” (AR), also known as NR3C4 (nuclear receptorsubfamily 3, group C, member 4), is a type of nuclear receptor that,when activated by binding an androgen receptor binder (e.g., anandrogenic hormone such as testosterone, or dihydrotestosterone) in thecytoplasm, is capable of translocating the androgenic hormone into thenucleus.

Exemplary androgen receptor-targeting epitopes which can be used in thecompounds described herein include, but are not limited to, an androgenreceptor agonist, a selective androgen-receptor modulator (SARM) (e.g.,enobosarm), an androgen receptor antagonist (e.g., bicalutamide,flutamide, nilutamide, or enzalutamide), a selective estrogen receptormodulator (SERM) (e.g., tamoxifen, toremifene, or raloxifene), anestrogen receptor antagonist (e.g., fulvestrant), a progestin (e.g.,megestrol acetate), an estrogen (e.g., estramustine), ketoconazole,abiraterone, darolutamide, or an analog thereof.

In certain embodiments, the nuclear steroid receptor-targeting epitopeis a selective androgen receptor modulator (SARM). In certainembodiments, the compound comprises at least one nuclear steroidreceptor-targeting epitope independently comprises an epitope derivedfrom testosterone, a testosterone ester (e.g., testosterone enanthate,propionate, cypionate, etc., or an analog thereof), enobosarm,BMS-564929, PS178990, LGD-4033 (ligandrol), LGD-2941, AC-262,356,JNJ-28330835, JNJ-37654032, JNJ-26146900, LGD-2226, LGD-3303,LGD-121071, LG-120907, S-40503, S-23, RAD-140, acetothiolutamide,andarine (S-4), LG-121071, TFM-4AS-1, YK-11, MK-0773 (PF-05314882),GSK2849466, GSK2881078, GSK8698, GSK4336, ACP-10.5, TT701, LY24.52473,1-(2-hydroxy-2-methyl-3-phenoxypropanoyl)-indoline-4-carbonitrile-derivatives(J Med Chem. 2014, 57(6), 2462-71), or an analog thereof.

In certain embodiments, a single atom on the nuclear receptor-targetingepitope as disclosed herein is replaced for attachment to the remainderof the compound (e.g., the moiety A-L- of Formula I and Formula II). Incertain embodiments, a halogen atom on a nuclear receptor-targetingepitope disclosed herein is replaced for attachment to the remainder ofthe compound. In certain embodiments, a hydrogen atom on a nuclearreceptor-targeting epitope disclosed herein is replaced for attachmentto the remainder of the compound. In certain embodiments, the hydrogenatom is on a heteroatom. In certain embodiments, the hydrogen atom is ona nitrogen. In certain embodiments, the hydrogen atom is on an oxygen.In certain embodiments, the hydrogen atom is on a carbon.

In certain embodiments, the moiety -B of Formula I or Formula IIcomprises at least one nuclear receptor-targeting epitope derived from:

or a stereoisomer or a mixture of stereoisomers thereof or an analogthereof.

These and other selective androgen receptor modulator (SARMs) which canbe used as a nuclear steroid receptor-targeting epitope in the compoundsdescribed herein can be found in U.S. Pat. Nos. 6,462,038, 6,777,427,WO2001/027086, WO2004/013104, WO2004/000816, WO2004/0113309,US2006/0211756, US2006/0063819, US2005/245485, US2005/250741,US2005/277681, WO2006/060108, WO2004/041277, WO2003/034987,US2006/0148893, US2006/0142387, WO2005/000795, WO2005/085185,WO2006/133216, WO2006/044707, WO2006/124447, WO2007/002181,WO2005/108351, WO2005/115361, and US2006/0160845.

In certain embodiments, the nuclear steroid receptor-targeting epitopeis a selective estrogen receptor modulator (SERM). In certainembodiments, the compound comprises at least one nuclear steroidreceptor-targeting epitope independently comprises an epitope derivedfrom anordrin, bazedoxifene, broparestrol (Acnestrol), clomifene(Clomid), cyclofenil (Sexovid), lasofoxifene (Fablyn), ormeloxifene(Centron, Novex, Novex-DS, Sevista), ospemifene (Osphena,deaminohydroxytoremifene), raloxifene (Evista), tamoxifen (Nolvadex),toremifene (Fareston; 4-chlorotamoxifen), acolbifene, afimoxifene(4-hydroxytamoxifen; metabolite of tamoxifen), elacestrant, enclomifene((E)-clomifene), endoxifen (4-hydroxy-N-desmethyltamoxifen; metaboliteof tamoxifen), zuclomifene ((Z)-clomifene), bazedoifene, arzoxifene,brilanestrant, clomifenoxide (clomiphene N-oxide; metabolite ofclomifene), droloxifene (3-hydroxytamoxifen), etacstil, fispemifene,GW-7604 (4-hydroxyetacstil), idoxifene (pyrrolidino-4-iodotamoxifen),levormeloxifene ((L)-ormeloxifene), miproxifene, nafoxidine, nitromifene(CI-628), panomifene, pipendoxifene (ERA-923), trioxifene, keoxifene,LY117018, onapristone, fareston (toremifine citrate) or zindoxifene(D-16726), or an analog thereof.

In certain embodiments, the SERM is classified structurally as atriphenylethylene (tamoxifen, clomifene, toremifene, droloxifene,idoxifene, ospemifene, fispemifene, afimoxifene, etc., or an analogthereof), a benzothiophene (raloxifene, arzoxifene, etc., or an analogthereof), an indole (bazedoxifene, zindoxifene, pipendoxifene, etc., oran analog thereof), a tetrahydronaphthalene (lasofoxifene, nafoxidine,etc., or an analog thereof), or a benzopyran (acolbifene, ormeloxifene,levormeloxifene, etc., or an analog thereof).

In certain embodiments, the nuclear steroid receptor-targeting epitopeis a selective estrogen receptor downregulator (SERD). In certainembodiments, the compound comprises at least one nuclear steroidreceptor-targeting epitope independently comprises an epitope derivedfrom fulvestrant, ARN-810, GW5638, or GW7604.

In certain embodiments, the nuclear steroid receptor-targeting epitopeis a selective progesterone receptor modulator (SPRM). In certainembodiments, the compound comprises at least one nuclear steroidreceptor-targeting epitope independently comprises an epitope derivedfrom ulipristal acetate, asoprisnil (J867), mifepristone, telapristone(CDB-4124, Proellex, Progenta), or an analog thereof.

In certain embodiments, the compound comprises at least one nuclearsteroid receptor-targeting epitope independently comprises an epitopederived from estrogen, estetrol, estriol, estrone, progesterone,enobosarm, bicalutamide, apalutamide, testosterone, dihydrotestosterone,estradiol, flutamide, nilutamide, enzalutamide, tamoxifen, toremifene,raloxifene, bazedoxifene, ospemifene, megestrol acetate, estramustine,abiraterone, LGD-2941, BMS-564929, ostarine, or an analog thereof.

In certain embodiments, at least one nuclear steroid receptor-targetingepitope is an androgen receptor-targeting epitope, and comprises:

or a stereoisomer or a mixture of stereoisomers thereof or an analogthereof, where the wavy line indicates the point of attachment to thelinking moiety or nuclear payload.

In certain embodiments, at least one nuclear steroid receptor-targetingepitope is an estrogen receptor-targeting epitope, and comprises:

or a stereoisomer or a mixture of stereoisomers thereof or an analogthereof, where the wavy line indicates the point of attachment to thelinking moiety or nuclear payload.

In certain embodiments, the nuclear steroid receptor-targeting epitopeis not, or does not contain, a peptide, protein, nanoparticle orantibody.

Exemplary compounds provided by the present disclosure include, but arenot limited to, a compound as shown in Table 1, or stereoisomer, mixtureof stereoisomers, hydrate, solvate, isotopically enriched analog orpharmaceutically acceptable salt thereof.

TABLE 1 No. Structure 1.1

1.2

1.3

ent. 1.3

1.4

1.5

1.6

1.7

1.8

1.9

1.10

1.11

1.12

1.13

1.14

1.15

1.16

1.17

1.18

1.19

1.20

1.21

1.22

1.23

1.24

1.25

1.26

1.27

1.28

1.29

1.30

1.31

1.32

1.33

1.34

1.35

1.36

1.37

1.38

1.39

1.40

1.41

1.42

1.43

1.44

1.45

1.46

1.47

1.48

1.49

1.50

1.51

1.52

1.53

1.54

1.55

1.56

1.57

1.58

1.59

1.60

1.61

1.62

1.63

1.64

1.65

1.66

1.67

1.68

1.69

1.70

1.71

1.72

1.73

1.74

1.75

1.76

1.77

1.78

1.79

1.80

1.81

1.82

1.83

1.84

1.85

1.86

1.87

1.88

1.89

1.90

1.91

1.92

1.93

1.94

1.95

1.96

1.97

1.98

1.99

1.101

1.102

1.103

1.104

1.105

1.106

1.107

1.108

1.109

1.110

1.111

1.112

1.113

1.114

1.115

1.116

1.117

1.118

1.119

1.120

1.121

1.122

1.123

1.124

1.125

1.126

1.127

1.128

1.129

1.130

1.131

1.132

1.133

1.134

1.135

1.136

1.137

1.138

1.139

1.140

1.141

1.142

1.143

1.144

1.145

1.146

1.147

1.148

1.149

1.150

1.151

1.152

1.153

1.154

1.155

1.156

1.157

1.158

1.159

1.160

1.161

1.162

1.163

1.164

1.165

1.166

1.167

1.168

1.169

1.170

1.171

_ 1.172

1.173

1.174

1.175

1.176

1.177

1.178

1.179

1.180

1.181

1.182

1.183

1.184

1.185

1.186

1.187

1.188

1.189

1.190

1.191

1.192

1.193

1.194

1.195

1.196

1.197

1.198

1.199

1.200

1.201

1.202

1.203

1.204

1.205

1.206

1.207

1.208

1.209

1.210

1.211

1.212

1.213

1.214

1.215

1.216

1.217

1.218

1.219

1.220

1.221

1.222

1.223

1.224

1.225

1.226

1.227

1.228

1.229

1.230

1.231

1.232

1.233

1.234

1.235

1.236

1.237

1.238

1.239

1.240

1.241

1.242

1.243

1.244

1.245

1.246

1.247

1.248

1.249

1.250

1.251

1.252

1.253

1.254

1.255

1.256

1.257

1.258

1.259

2.1

2.2

2.3

2.4

2.5

2.6

2.7

2.8

2.9

2.10

2.11

2.12

2.13

2.14

2.15

2.16

2.17

2.18

2.19

2.20

2.21

2.22

2.23

2.24

2.25

2.26

2.27

2.28

2.29

2.30

2.31

2.32

2.33

2.34

2.35

2.36

2.37

2.38

2.39

2.40

2.41

2.42

2.43

2.44

2.45

2.46

2.47

3.1

3.2

3.3

3.4

3.5

3.6

3.7

3.8

3.9

3.10

3.11

3.12

3.13

3.14

3.15

3.16

3.17

3.18

3.19

3.20

3.21

3.22

3.27

3.29

3.31

3.32

3.33

3.34

3.35

3.36

3.37

3.38

3.39

3.40

3.41

3.42

3.43

3.44

3.45

3.46

3.47

3.48

3.49

3.50

3.51

3.52

3.53

3.54

3.61

3.62

3.63

3.64

3.65

3.66

3.67

3.68

3.69

3.70

3.71

3.72

3.73

3.74

3.75

3.76

3.77

3.78

3.85

3.86

3.87

3.88

3.89

3.90

3.91

3.92

3.93

3.94

3.95

3.96

3.97

3.98

3.99

3.100

3.101

3.102

3.103

3.104

3.105

3.106

3.107

3.108

3.109

3.110

3.111

3.112

3.113

3.114

3.115

3.116

3.117

3.118

3.119

3.120

3.121

3.122

3.123

3.124

3.125

3.126

3.127

3.128

3.129

3.130

3.131

3.132

3.133

3.134

3.135

3.136

3.137

3.138

3.139

3.140

3.141

3.142

3.143

3.144

3.145

3.146

3.147

3.148

3.149

3.150

3.151

3.152

3.153

3.154

3.155

3.156

3.157

3.158

3.159

3.160

3.161

3.162

3.163

3.164

3.165

3.166

3.167

3.168

3.169

3.170

3.171

3.172

3.173

3.174

3.175

3.176

3.177

3.178

3.179

3.180

3.181

3.182

3.183

3.184

3.185

3.186

3.187

3.188

3.189

3.190

3.191

3.192

3.193

3.194

3.195

3.196

3.197

3.198

3.199

3.200

3.201

3.202

3.203

3.204

3.205

3.206

3.207

3.208

3.209

3.210

3.211

3.212

3.213

3.214

3.215

3.216

3.217

3.218

3.219

3.220

3.221

3.222

3.223

3.224

3.225

3.226

3.227

3.228

3.229

3.230

3.231

3.232

3.233

3.234

3.235

3.236

3.237

3.238

3.239

3.240

3.241

3.242

3.243

3.244

3.245

3.246

3.247

3.248

3.249

3.250

3.251

3.252

3.253

3.254

3.255

3.256

3.257

3.258

3.259

3.260

3.261

3.262

3.263

3.264

Methods of Treatment

Provided herein are compounds which can be used to treat, prevent,and/or delay the onset and/or development of cancer. Accordingly, incertain embodiments, provided is a method for the treatment of cancer,comprising administering to a subject in need of treatment atherapeutically-effective amount of a compound or composition describedherein. Certain embodiments provide a method of potentiation ofcytotoxic cancer therapy in a subject in recognized need of suchtreatment comprising administering to the subject a therapeuticallyacceptable amount of a compound or composition described herein.

It is contemplated that a patient having any cancer may benefit frombeing treated with the compounds and compositions described herein.Accordingly, in certain embodiments, the cancer is liver cancer,melanoma, Hodgkin's disease, non-Hodgkin's lymphomas, acute lymphocyticleukemia, chronic lymphocytic leukemia, multiple myeloma, neuroblastoma,breast carcinoma, ovarian carcinoma, lung carcinoma, Wilms' tumor,cervical carcinoma, testicular carcinoma, soft-tissue sarcoma, chroniclymphocytic leukemia, primary macroglobulinemia, bladder carcinoma,chronic granulocytic leukemia, primary brain carcinoma, malignantmelanoma, small-cell lung carcinoma, stomach carcinoma, colon carcinoma,malignant pancreatic insulinoma, malignant carcinoid carcinoma,malignant melanoma, choriocarcinoma, mycosis fungoide, head neckcarcinoma, osteogenic sarcoma, pancreatic carcinoma, acute granulocyticleukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma,genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma,malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma,endometrial carcinoma, polycythemia vera, essential thrombocytosis,adrenal cortex carcinoma, skin cancer, or prostatic carcinoma. Incertain embodiments, the cancer is bladder cancer, a blood cancer, suchas leukemia (e.g., chronic leukemia, chronic lymphocytic leukemia (CLL,etc.) or lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, lowgrade lymphoma, high grade lymphoma), lung cancer (e.g., small cell lungcancer), breast cancer, fallopian tube cancer, glioblastoma multiforme,head and neck cancer, esophageal cancer, ovarian cancer, pancreaticcancer, peritoneal cancer, prostate cancer, testicular cancer, skincancer (e.g., melanoma) or uterine cancer. In certain embodiments, thecancer is bladder cancer, breast cancer, fallopian tube cancer, ovariancancer, prostate cancer, peritoneal cancer, testicular cancer,endometrial cancer, or uterine cancer.

In certain embodiments, the compounds and compositions as describedherein are tailored to target cancers which overexpress a specificreceptor, such as, but not limited to, androgen receptors, estrogenreceptors, progesterone receptors, and/or glucocorticoid receptors byincluding an epitope which targets that specific nuclear receptor. Theepitope can be derived from a steroid hormone or any non-steroidal drugwhich targets that particular receptor.

In certain embodiments, provided is a method of treating or preventingan androgen receptor overexpressing cancer, comprising administering aneffective amount of a compound, or a pharmaceutically acceptable salt ofsolvate thereof, comprising at least one nuclear payload and at leastone androgen receptor-targeting epitope to an individual in needthereof. Specific cancers which are contemplated to be treated by suchmethods include, but are not limited to, prostate, breast, triplenegative breast cancer, bladder, or liver cancer. Also provided is amethod of treating or preventing metastatic castration-resistantprostate cancer (mCRPC), comprising administering an effective amount ofa compound or composition as described herein, or a pharmaceuticallyacceptable salt or solvate thereof, to an individual in need thereof.

In certain embodiments, provided is a method of treating or preventingan androgen receptor overexpressing cancer, comprising administering aneffective amount of a compound, or a pharmaceutically acceptable salt orsolvate thereof, comprising at least one nuclear payload and at leastone androgen receptor-targeting epitope to an individual in needthereof. In certain embodiments, the cancer is prostate, breast, triplenegative breast cancer, bladder, or liver cancer. In certainembodiments, the androgen receptor-targeting epitope comprises anandrogen receptor agonist, a selective androgen-receptor modulator(SARM), an androgen receptor antagonist, a selective estrogen receptormodulator (SERM), an estrogen receptor antagonist, a progestin, or anestrogen. In certain embodiments, the androgen receptor-targetingepitope comprises enobosarm, bicalutamide, flutamide, nilutamide,enzalutamide, tamoxifen, toremifene, raloxifene, fulvestrant, megestrolacetate, estramustine, ketoconazole, abiraterone, darolutamide, or ananalog thereof. In certain embodiments, the androgen receptor-targetingepitope comprises enobosarm, bicalutamide, flutamide, nilutamide,enzalutamide, tamoxifen, toremifene, raloxifene, fulvestrant, megestrolacetate, estramustine, ketoconazole, abiraterone, or an analog thereof.In certain embodiments, the nuclear payload comprises a PARP inhibitor.

In certain embodiments, provided is a method of treating or preventingan estrogen and/or progesterone receptor overexpressing cancer,comprising administering an effective amount of a compound, or apharmaceutically acceptable salt or solvate thereof, comprising at leastone nuclear payload and at least one estrogen and/or progesteronereceptor-targeting epitope to an individual in need thereof. Specificcancers which are contemplated to be treated by such methods include,but are not limited to, breast, uterine, or ovarian cancer.

In certain embodiments, provided is a method of treating or preventing aglucocorticoid receptor overexpressing cancer, comprising administeringan effective amount of a compound, or a pharmaceutically acceptable saltor solvate thereof, comprising at least one nuclear payload and at leastone glucocorticoid receptor-targeting epitope to an individual in needthereof. Specific cancers which are contemplated to be treated by suchmethods include, but are not limited to, breast, uterine, or ovariancancer. Specific cancers which are contemplated to be treated by suchmethods include, but are not limited to, prostate, possibly breast,uterine, ovarian.

Breast cancer includes ductal carcinoma in situ (DCIS) and invasivebreast cancer. Breast cancers can occur in milk ducts, milk-producinglobules and connective tissues. Breast cancer includes estrogen receptor(ER) negative and hormone receptor (HR) negative, and also can becategorized as Group 3 (HER-2 positive) or Group 4 (basal-like).

Prostate cancer is a cancer which develops in the prostate, a gland inthe male reproductive system. It occurs when cells of the prostatemutate and begin to multiply uncontrollably. These cells may metastasize(metastatic prostate cancer) from the prostate to virtually any otherpart of the body, particularly the bones and lymph nodes, but thekidney, bladder and even the brain, among other tissues. Prostate cancermay cause pain, difficulty in urinating, problems during sexualintercourse, erectile dysfunction. Other symptoms can potentiallydevelop during later stages of the disease. Rates of detection ofprostate cancers vary widely across the world, with South and East Asiadetecting less frequently than in Europe, and especially the UnitedStates. Prostate cancer develops most frequently in men over the age offifty and is one of the most prevalent types of cancer in men. However,many men who develop prostate cancer never have symptoms, undergo notherapy, and eventually die of other causes. This is because cancer ofthe prostate is, in most cases, slow-growing, and because most of thoseaffected are over the age of 60. Hence, they often die of causesunrelated to prostate cancer. Many factors, including genetics and diet,have been implicated in the development of prostate cancer. The presenceof prostate cancer may be indicated by symptoms, physical examination,prostate specific antigen (PSA), or biopsy. There is concern about theaccuracy of the PSA test and its usefulness in screening. Suspectedprostate cancer is typically confirmed by taking a biopsy of theprostate and examining it under a microscope. Further tests, such as CTscans and bone scans, may be performed to determine whether prostatecancer has spread. Combination with primarily surgery and radiationtherapy, or other treatments such as hormonal therapy, chemotherapy,proton therapy, cryosurgery, high intensity focused ultrasound (HIFU)are also contemplated.

Certain embodiments provide a method of inhibiting PARP in a subject inrecognized need of such treatment comprising administering to thesubject a therapeutically acceptable amount of a compound or compositiondescribed herein. In one embodiment, provided herein is a method oftreating a disease ameliorated by the inhibition of PARP comprisingadministering to a subject in need of treatment atherapeutically-effective amount of a compound or composition describedherein.

Certain embodiments provide a method of treating leukemia, colon cancer,glioblastomas, lymphomas, melanomas, carcinomas of breast, or cervicalcarcinomas in a subject in recognized need of such treatment comprisingadministering to the subject a therapeutically acceptable amount of acompound or composition described herein.

In some embodiments, provided herein is a method of treatment of acancer deficient in Homologous Recombination (HR) dependent DNA doublestrand break (DSB) repair pathway, which includes administering to asubject in need of treatment a therapeutically-effective amount of acompound of composition described herein. In certain embodiments, thecancer includes one or more cancer cells having a reduced or abrogatedability to repair DNA DSB by HR relative to normal cells. In someembodiments, the cancer cells have a BRCA1 or BRCA2 deficient phenotype.In some embodiments, the cancer cells are deficient in BRCA1 or BRCA2.In some embodiments, the methods provided herein involve treatment of anindividual who is heterozygous for a mutation in a gene encoding acomponent of the HR dependent DNA DSB repair pathway. In certainembodiment, the individual is heterozygous for a mutation in BRCA1and/or BRCA2. In some embodiments, the method of treatment of a cancerincludes treatment of breast, ovary, pancreas and/or prostate cancer. Insome embodiments, the method of treatment of a cancer further includesadministration of ionizing radiation or a chemotherapeutic agent.

The primary function of the DNA mismatch repair (MMR) system is toeliminate single-base mismatches and insertion-deletion loops that mayarise during DNA replication. Insertion-deletion loops result from gainsor losses of short repeat units within microsatellite sequences, alsoknown as microsatellite instability (MSI). At least six different MMRproteins are required. For mismatch recognition, the MSH2 protein formsa heterodimer with either MSH6 or MSH3 depending on the type of lesionto be repaired (MSH6 is required for the correction of single-basemispairs, whereas both MSH3 and MSH6 may contribute to the correction ofinsertion-deletion loops). A heterodimer of MLH1 and PMS2 coordinatesthe interplay between the mismatch recognition complex and otherproteins necessary for MMR. These additional proteins may include atleast exonuclease 1 (EXO 1), possibly helicase(s), proliferating cellnuclear antigen (PCNA), single-stranded DNA-binding protein (RPA), andDNA polymerases δ and ε. In addition to PMS2, MLH1 may heterodimerizewith two additional proteins, MLH3 and PMS1. Recent observationsindicate that PMS2 is required for the correction of single-basemismatches, and PMS2 and MLH3 both contribute to the correction ofinsertion-deletion loops. Additional homologs of the human MMR proteinsare known that are required for functions other than MMR. These proteinsinclude MSH4 and MSH5 that are necessary for meiotic (and possiblymitotic) recombination but are not presumed to participate in MMR.

Germline mutations of human MMR genes cause susceptibility to hereditarynonpolyposis colon cancer (HNPCC), one of the most common cancersyndromes in humans. An excess of colon cancer and a defined spectrum ofextracolonic cancers, diagnosed at an early age and transmitted as anautosomal dominant trait, constitute the clinical definition of thesyndrome. MSI, the hallmark of HNPCC, occurs in approximately 15% to 25%of sporadic tumors of the colorectum and other organs as well. Accordingto international criteria, a high degree of MSI (MSI-H) is defined asinstability at two or more of five loci or >30% to 40% of allmicrosatellite loci studied, whereas instability at fewer loci isreferred to as MSI-low (MSI-L). MSI occurs in a substantial proportion(2% to 50% of tumors) among non-HNPCC cancers (e.g., cancers of thebreast, prostate, and lung). On the basis of the proportion of unstablemarkers, categories MSS, MSI-L, and MSI-H can be distinguished in thesecancers in analogy to HNPCC cancers. In one embodiment is a method fortreating a cancer deficient in mismatch DNA repair pathway. In anotherembodiment is a method for treating a cancer demonstratingmicrosatellite instability due to reduced or impaired DNA repairpathways. In another embodiment is a method for treating a cancerdemonstrating genomic instability due to reduced or impaired DNA repairpathways.

In certain embodiments, a compound or composition described herein, maybe used in the preparation of a medicament for the treatment of cancerwhich is deficient in Homologous Recombination (HR) dependent DNA doublestrand break (DSB) repair activity, or in the treatment of a patientwith a cancer which is deficient in HR dependent DNA DSB repairactivity, which includes administering to said patient atherapeutically-effective amount of the compound or composition.

The HR dependent DNA DSB repair pathway repairs double-strand breaks(DSBs) in DNA via homologous mechanisms to reform a continuous DNAhelix. The components of the HR dependent DNA DSB repair pathwayinclude, but are not limited to, ATM (NM_000051), RAD51 (NM_002875),RAD51L1 (NM_002877), RAD51C (NM_002876), RAD51L3 (NM_002878), DMC1(NM_007068), XRCC2 (NM_005431), XRCC3 (NM_005432), RAD52 (NM_002879),RAD54L (NM_003579), RAD54B (NM_012415), BRCA1 (NM_007295), BRCA2(NM_000059), RAD50 (NM_005732), MRE1 IA (NM_005590) and NBS11M_00248_5). Other proteins involved in the HR dependent DNA DSB repairpathway include regulatory factors such as EMSY (Wood, et al., Science,291, 1284-1289 (2001); Khanna et al., Nat. Genet. 27(3): 247-254 (2001);and Hughes-Davies, et al., Cell, 115, pp 523-535).

In some embodiments, a cancer which is deficient in HR dependent DNA DSBrepair includes one or more cancer cells which have a reduced orabrogated ability to repair DNA DSBs through that pathway, relative tonormal cells, i.e. the activity of the HR dependent DNA DSB repairpathway are reduced or abolished in the one or more cancer cells.

In certain embodiments, the activity of one or more components of the HRdependent DNA DSB repair pathway is abolished in the one or more cancercells of an individual having a cancer which is deficient in HRdependent DNA DSB repair. Components of the HR dependent DNA DSB repairpathway include the components listed above.

In some embodiments, the cancer cells have a BRCA1 and/or a BRCA2deficient phenotype, i.e., BRCA1 and/or BRCA2 activity is reduced orabolished in the cancer cells. In certain embodiments, cancer cells withthis phenotype are deficient in BRCA1 and/or BRCA2, i.e., expressionand/or activity of BRCA1 and/or BRCA2 is reduced or abolished in thecancer cells, for example by means of mutation or polymorphism in theencoding nucleic acid, or by means of amplification, mutation orpolymorphism in a gene encoding a regulatory factor, for example theEMSY gene which encodes a BRCA2 regulatory factor or by an epigeneticmechanism such as gene promoter methylation.

BRCA1 and BRCA2 are tumor suppressors whose wild-type alleles arefrequently lost in tumors of heterozygous carriers. BRCA1 and/or BRCA2mutations are associated with breast cancer. Amplification of the EMSYgene, which encodes a BRCA2 binding factor, is associated with breastand ovarian cancer (Jasin M., Oncogene, 21(58), 8981-93 (2002); Tutt, etal, Trends Mol. Med., 8(12), 571-6, (2002); and Radice, P. J., Exp ClinCancer Res., 21(3 Suppl), 9-12 (2002)).

Carriers of mutations in BRCA1 and/or BRCA2 are also at elevated risk ofcancer of the ovary, prostate and pancreas.

In some embodiments, the individual is heterozygous for one or morevariations, such as mutations and polymorphisms, in BRCA1 and/or BRCA2or a regulator thereof. The detection of variation in BRCA1 and BRCA2 isdescribed, for example in EP 699 754, EP 705 903, Neuhausen, S. L. andOstrander, E. A., Genet. Test, 1, 75-83 (1992); Janatova M., et al,Neoplasma, 50(4), 246-50 (2003). Determination of amplification of theBRCA2 binding factor EMSY is described in Hughes-Davies, et al., Cell,115, 523-535.

In certain instances, mutations and polymorphisms associated with cancerare detected at the nucleic acid level by detecting the presence of avariant nucleic acid sequence or at the protein level by detecting thepresence of a variant (i.e. a mutant or allelic variant) polypeptide.

In certain embodiments, it is contemplated that the compounds describedherein are useful for patients who have relapsed or become refractory.The term “relapsed” refers to disease (or cancer) that reappears orgrows again after a period of remission. The term “refractory” is usedto describe when the cancer does not respond to treatment or when theresponse to treatment does not last very long. For example, thecompounds herein may be useful for treating cancer in patients who havepreviously been treated with the cancer therapies described herein, e.g.enzalutamide.

Compositions

Compositions, including pharmaceutical compositions, of any of thecompounds detailed herein are embraced by this disclosure. Thus,provided herein are pharmaceutical compositions comprising a compound ofthe disclosure, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier or excipient. The pharmaceuticalcompositions provided herein may take a form suitable for oral, buccal,parenteral (e.g., intravenous, intramuscular, infusion or subcutaneousinjection), nasal, topical or rectal administration, or a form suitablefor administration by inhalation.

A compound as described herein may, in one aspect, be in a purifiedform. Compositions comprising a compound as described herein, or a saltthereof, are provided, such as compositions of substantially purecompounds. In some embodiments, a composition comprising a compound asdescribed herein, or a salt thereof, is in substantially pure form.Unless otherwise stated, “substantially pure” refers to a compositionwhich contains no more than 35% impurity, wherein the impurity denotes acompound other than the desired compound, or a salt thereof, whichcomprises the majority of the composition. In one variation, acomposition of substantially pure compound, or a salt thereof, isprovided wherein the composition contains no more than 25% impurity. Inanother variation, a composition of substantially pure compound, or asalt thereof, is provided wherein the composition contains or no morethan 20% impurity. In still another variation, a composition ofsubstantially pure compound, or a salt thereof, is provided wherein thecomposition contains or no more than 10% impurity. In a furthervariation, a composition of substantially pure compound, or a saltthereof, is provided wherein the composition contains or no more than 5%impurity. In another variation, a composition of substantially purecompound, or a salt thereof, is provided wherein the compositioncontains or no more than 3% impurity. In still another variation, acomposition of substantially pure compound, or a salt thereof, isprovided wherein the composition contains or no more than 1% impurity.In a further variation, a composition of substantially pure compound, ora salt thereof, is provided wherein the composition contains or no morethan 0.5% impurity.

In certain embodiments, pharmaceutical compositions are formulated inany manner, including using one or more physiologically acceptablecarriers comprising excipients and/or auxiliaries which facilitateprocessing of the active compounds into pharmaceutical compositions. Insome embodiments, proper formulation is dependent upon the route ofadministration chosen. In various embodiments, any techniques, carriers,and excipients are used as suitable.

Provided herein are pharmaceutical compositions that include a compounddescribed herein and a pharmaceutically acceptable diluent(s),excipient(s), and/or carrier(s). In addition, in some embodiments, thecompounds described herein are administered as pharmaceuticalcompositions in which compounds described herein are mixed with otheractive ingredients, as in combination therapy.

A pharmaceutical composition, as used herein, refers to a mixture of acompound described herein with other chemical components, such ascarriers, stabilizers, diluents, dispersing agents, suspending agents,thickening agents, and/or excipients. In certain embodiments, apharmaceutical composition facilitates administration of the compound toan organism. In some embodiments, practicing the methods of treatment oruse provided herein, includes administering or using a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundprovided herein. In specific embodiments, the methods of treatmentprovided for herein include administering such a pharmaceuticalcomposition to a mammal having a disease or condition to be treated. Inone embodiment, the mammal is a human. In some embodiments, thetherapeutically effective amount varies widely depending on the severityof the disease, the age and relative health of the subject, the potencyof the compound used and other factors. In various embodiments, thecompounds described herein are used singly or in combination with one ormore therapeutic agents as components of mixtures.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated for intravenous injections. In certain aspects, theintravenous injection formulations provided herein are formulated asaqueous solutions, and, in some embodiments, in physiologicallycompatible buffers such as Hank's solution, Ringer's solution, orphysiological saline buffer. In certain embodiments, the pharmaceuticalcompositions provided herein are formulated for transmucosaladministration. In some aspects, transmucosal formulations includepenetrants appropriate to the barrier to be permeated. In certainembodiments, the pharmaceutical compositions provided herein areformulated for other parenteral injections, appropriate formulationsinclude aqueous or nonaqueous solutions, and in one embodiment, withphysiologically compatible buffers or excipients.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated for oral administration. In certain aspects, the oralformulations provided herein comprise compounds described herein thatare formulated with pharmaceutically acceptable carriers or excipients.Such carriers enable the compounds described herein to be formulated astablets, powders, pills, dragees, capsules, liquids, gels, syrups,elixirs, slurries, suspensions and the like, for oral ingestion by apatient to be treated.

In some embodiments, pharmaceutical compositions for oral use areobtained by mixing one or more solid excipient with one or more of thecompounds described herein, optionally grinding the resulting mixture,and processing the mixture of granules, after adding suitableauxiliaries, if desired, to obtain tablets or dragee cores. Suitableexcipients include, in particular, fillers such as sugars, includinglactose, sucrose, mannitol, or sorbitol; cellulose preparations such as:for example, maize starch, wheat starch, rice starch, potato starch,gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or otherssuch as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. Ifdesired, disintegrating agents are optionally added, such as thecross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, oralginic acid or a salt thereof such as sodium alginate.

In certain embodiments, provided herein is a pharmaceutical compositionformulated as dragee cores with suitable coatings. In certainembodiments, concentrated sugar solutions are used in forming thesuitable coating, and optionally contain gum arabic, talc,polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titaniumdioxide, lacquer solutions, and suitable organic solvents or solventmixtures. In some embodiments, dyestuffs and/or pigments are added totablets, dragees and/or the coatings thereof for, e.g., identificationor to characterize different combinations of active compound doses.

In certain embodiments, pharmaceutical compositions which are usedinclude orally include push-fit capsules made of gelatin, as well assoft, sealed capsules made of gelatin and a plasticizer, such asglycerol or sorbitol. In some embodiments, the push-fit capsules containthe active ingredients in admixture with filler such as lactose, binderssuch as starches, and/or lubricants such as talc or magnesium stearateand, optionally, stabilizers. In certain embodiments, in soft capsules,the active compounds are dissolved or suspended in suitable liquids,such as fatty oils, liquid paraffin, or liquid polyethylene glycols. Inaddition, stabilizers are optionally added. In certain embodiments, theformulations for oral administration are in dosages suitable for suchadministration.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated for buccal or sublingual administration. In certainembodiments, buccal or sublingual compositions take the form of tablets,lozenges, or gels formulated in a conventional manner. In certainembodiments, parenteral injections involve bolus injection or continuousinfusion. In some embodiments, formulations for injection are presentedin unit dosage form, e.g., in ampoules or in multi-dose containers, withan added preservative. In some embodiments, the pharmaceuticalcomposition described herein is in a form suitable for parenteralinjection as a sterile suspensions, solutions or emulsions in oily oraqueous vehicles, and optionally contains formulatory agents such assuspending, stabilizing and/or dispersing agents. Pharmaceuticalformulations for parenteral administration include aqueous solutions ofthe active compounds in water-soluble form. In some embodiments,suspensions of the active compounds are prepared as appropriate oilyinjection suspensions. Suitable lipophilic solvents or vehicles includefatty oils such as sesame oil, or synthetic fatty acid esters, such asethyl oleate or triglycerides, or liposomes. In certain embodiments,aqueous injection suspensions contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspensions also contain suitablestabilizers or agents which increase the solubility of the compounds toallow for the preparation of highly concentrated solutions. Inalternative embodiments, the active ingredient is in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

In some embodiments, the compounds described herein are administeredtopically. In specific embodiments, the compounds described herein areformulated into a variety of topically administrable compositions, suchas solutions, suspensions, lotions, gels, pastes, medicated sticks,balms, creams or ointments. Such pharmaceutical compounds optionallycontain solubilizers, stabilizers, tonicity enhancing agents, buffersand/or preservatives.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated for transdermal administration of compounds describedherein. In some embodiments, administration of such compositions employstransdermal delivery devices and transdermal delivery patches. Incertain embodiments, the compositions are lipophilic emulsions orbuffered, aqueous solutions, dissolved and/or dispersed in a polymer oran adhesive. Such patches include those constructed for continuous,pulsatile, or on demand delivery of pharmaceutical agents. In someembodiments, transdermal delivery of the compounds described herein isaccomplished by use of iontophoretic patches and the like. In certainembodiments, the rate of absorption is slowed by using rate-controllingmembranes or by trapping the compound within a polymer matrix or gel.Conversely, absorption enhancers are optionally used to increaseabsorption. Absorption enhancer and carrier include absorbablepharmaceutically acceptable solvents that assist in passage of thecompound through the skin. For example, transdermal devices are in theform of a bandage comprising a backing member, a reservoir containingthe compound optionally with carriers, optionally a rate controllingbarrier to deliver the compound to the skin of the host at a controlledand predetermined rate over a prolonged period of time, and means tosecure the device to the skin.

In certain embodiments, the pharmaceutical compositions provided hereinare formulated for administration by inhalation. In certain embodiments,in such pharmaceutical compositions formulated for inhalation, thecompounds described herein are in a form as an aerosol, a mist or apowder. In some embodiments, pharmaceutical compositions describedherein are conveniently delivered in the form of an aerosol spraypresentation from pressurized packs or a nebulizer, with the use of asuitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In certain aspects of a pressurized aerosol, thedosage unit is determined by providing a valve to deliver a meteredamount. In certain embodiments, capsules and cartridges of, such as, byway of example only, gelatin for use in an inhaler or insufflator isformulated containing a powder mix of the compound described herein anda suitable powder base such as lactose or starch.

In some embodiments, the compounds described herein are formulated inrectal compositions such as enemas, rectal gels, rectal foams, rectalaerosols, suppositories, jelly suppositories, or retention enemas. Incertain embodiments, rectal compositions optionally contain conventionalsuppository bases such as cocoa butter or other glycerides, as well assynthetic polymers such as polyvinylpyrrolidone, PEG, and the like. Incertain suppository forms of the compositions, a low-melting wax suchas, but not limited to, a mixture of fatty acid glycerides, optionallyin combination with cocoa butter is first melted.

In various embodiments provided herein, the pharmaceutical compositionsare formulated in a conventional manner using one or morephysiologically acceptable carriers comprising excipients andauxiliaries which facilitate processing of the active compounds intopharmaceutically acceptable preparations. In certain embodiments, properformulation is dependent upon the route of administration chosen. Invarious embodiments, any of the techniques, carriers, and excipients isused as suitable. In some embodiments, pharmaceutical compositionscomprising a compound described herein are manufactured in aconventional manner, such as, by way of example only, by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or compression processes.

In certain embodiments, the pharmaceutical compositions include at leastone pharmaceutically acceptable carrier, diluent or excipient and acompound described herein described herein as an active ingredient infree-acid or free-base form, or in a pharmaceutically acceptable saltform. In addition, the methods and pharmaceutical compositions describedherein include the use of N-oxides, crystalline forms (also known aspolymorphs), as well as active metabolites of these compounds having thesame type of activity. In some situations, compounds described hereinexist as tautomers. All tautomers are included within the scope of thecompounds presented herein. Additionally, included herein are thesolvated and unsolvated forms of the compounds described herein.Solvated compounds include those that are solvated with pharmaceuticallyacceptable solvents such as water, ethanol, and the like. The solvatedforms of the compounds presented herein are also considered to bedisclosed herein. In some embodiments, the pharmaceutical compositionsdescribed herein include other medicinal or pharmaceutical agents,carriers, adjuvants, such as preserving, stabilizing, wetting oremulsifying agents, solution promoters, salts for regulating the osmoticpressure, and/or buffers. In additional embodiments, the pharmaceuticalcompositions described herein also contain other therapeuticallyvaluable substances.

Methods for the preparation of compositions containing the compoundsdescribed herein include formulating the compounds with one or moreinert, pharmaceutically acceptable excipients or carriers to form asolid, semi-solid or liquid. Solid compositions include, but are notlimited to, powders, tablets, dispersible granules, capsules, cachets,and suppositories. Liquid compositions include solutions in which acompound is dissolved, emulsions comprising a compound, or a solutioncontaining liposomes, micelles, or nanoparticles comprising a compoundas disclosed herein. Semi-solid compositions include, but are notlimited to, gels, suspensions and creams. In various embodiments, thecompositions are in liquid solutions or suspensions, solid formssuitable for solution or suspension in a liquid prior to use, or asemulsions. These compositions optionally contain minor amounts ofnontoxic, auxiliary substances, such as wetting or emulsifying agents,pH buffering agents, and so forth.

In some embodiments, a composition comprising a compound describedherein takes the form of a liquid where the agents are present insolution, in suspension or both. In some embodiments, when thecomposition is administered as a solution or suspension a first portionof the agent is present in solution and a second portion of the agent ispresent in particulate form, in suspension in a liquid matrix. In someembodiments, a liquid composition includes a gel formulation. In otherembodiments, the liquid composition is aqueous.

Useful aqueous suspension optionally contain one or more polymers assuspending agents. Useful polymers include water-soluble polymers suchas cellulosic polymers, e.g., hydroxypropyl methylcellulose, andwater-insoluble polymers such as cross-linked carboxyl-containingpolymers. Useful compositions optionally comprise an mucoadhesivepolymer, selected for example from carboxymethylcellulose, carbomer(acrylic acid polymer), poly(methylmethacrylate), polyacrylamide,polycarbophil, acrylic acid/butyl acrylate copolymer, sodium alginateand dextran.

Useful compositions optionally include solubilizing agents to aid in thesolubility of a compound described herein. The term “solubilizing agent”generally includes agents that result in formation of a micellarsolution or a true solution of the agent. Solubilizing agents includecertain acceptable nonionic surfactants, for example polysorbate 80, andophthalmically acceptable glycols, polyglycols, e.g., polyethyleneglycol 400, and glycol ethers.

Useful compositions optionally include one or more pH adjusting agentsor buffering agents, including acids such as acetic, boric, citric,lactic, phosphoric and hydrochloric acids; bases such as sodiumhydroxide, sodium phosphate, sodium borate, sodium citrate, sodiumacetate, sodium lactate and tris-hydroxymethylaminomethane; and bufferssuch as citrate/dextrose, sodium bicarbonate and ammonium chloride. Suchacids, bases and buffers are included in an amount required to maintainpH of the composition in an acceptable range.

Useful compositions optionally include one or more salts in an amountrequired to bring osmolality of the composition into an acceptablerange. Such salts include those having sodium, potassium or ammoniumcations and chloride, citrate, ascorbate, borate, phosphate,bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable saltsinclude sodium chloride, potassium chloride, sodium thiosulfate, sodiumbisulfite and ammonium sulfate.

Certain useful compositions optionally include one or more preservativesto inhibit microbial activity. Suitable preservatives includemercury-containing substances such as merfen and thiomersal; stabilizedchlorine dioxide; and quaternary ammonium compounds such as benzalkoniumchloride, cetyltrimethylammonium bromide and cetylpyridinium chloride.

Some useful compositions optionally include one or more surfactants toenhance physical stability or for other purposes. Suitable nonionicsurfactants include polyoxyethylene fatty acid glycerides and vegetableoils, e.g., polyoxyethylene (60) hydrogenated castor oil; andpolyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10,octoxynol 40.

Certain useful compositions optionally one or more antioxidants toenhance chemical stability where required. Suitable antioxidantsinclude, by way of example only, ascorbic acid and sodium metabisulfite.

In some embodiments, aqueous suspension compositions are packaged insingle-dose non-reclosable containers. In alternative embodiments,multiple-dose reclosable containers are used, in which case it istypical to include a preservative in the composition.

In various embodiments, any delivery system for hydrophobicpharmaceutical compounds is employed. Liposomes and emulsions areexamples of delivery vehicles or carriers for hydrophobic drugs. Incertain embodiments, certain organic solvents such asN-methylpyrrolidone are employed. In some embodiments, the compounds aredelivered using a sustained-release system, such as semipermeablematrices of solid hydrophobic polymers containing the therapeutic agent.Various sustained-release materials are utilized in the embodimentsherein. In certain embodiments, sustained-release capsules release thecompounds for a few weeks up to over 100 days. In some embodiments,depending on the chemical nature and the biological stability of thetherapeutic reagent, additional strategies for protein stabilization areemployed.

In certain embodiments, the formulations or compositions describedherein benefit from and/or optionally comprise antioxidants, metalchelating agents, thiol containing compounds and other generalstabilizing agents. Examples of such stabilizing agents, include, butare not limited to: (a) about 0.5% to about 2% w/v glycerol, (b) about0.1% to about 1% w/v methionine, (c) about 0.1% to about 2% w/vmonothioglycerol, (d) about 1 mM to about 10 mM EDTA, (c) about 0.01% toabout 2% w/v ascorbic acid, (f) 0.003% to about 0.02% w/v polysorbate80, (g) 0.001% to about 0.05% w/v. polysorbate 20, (h) arginine, (i)heparin, (j) dextran sulfate, (k) cyclodextrins, (1) pentosanpolysulfate and other heparinoids, (m) divalent cations such asmagnesium and zinc; or (n) combinations thereof.

Dosing and Treatment Regimens

In certain embodiments, the compositions containing the compound(s)described herein are administered for prophylactic and/or therapeutictreatments. In certain therapeutic applications, the compositions areadministered to a patient already suffering from a disease or condition,in an amount sufficient to cure or at least partially arrest thesymptoms of the disease or condition. In some embodiments, amountseffective for this use will depend on the severity and course of thedisease or condition, previous therapy, the patient's health status,weight, and response to the drugs, and the judgment of the treatingphysician. In certain instances, it is considered appropriate for thecaregiver to determine such therapeutically effective amounts by routineexperimentation (including, but not limited to, a dose escalationclinical trial).

In certain prophylactic applications, compositions containing thecompounds described herein are administered to a patient susceptible toor otherwise at risk of a particular disease, disorder or condition. Insome embodiments, the amount administered is defined to be a“prophylactically effective amount or dose.” In certain embodiments ofthis use, the precise amounts of compound administered depend on thepatient's state of health, weight, and the like. In some embodiments, itis considered appropriate for the caregiver to determine suchprophylactically effective amounts by routine experimentation (e.g., adose escalation clinical trial). In certain embodiments, when used in apatient, effective amounts for this use will depend on the severity andcourse of the disease, disorder or condition, previous therapy, thepatient's health status and response to the drugs, and the judgment ofthe treating physician.

In certain instances, a patient's condition does not improve or does notsignificantly improve following administration of a compound orcomposition described herein and, upon the doctor's discretion theadministration of the compounds is optionally administered chronically,that is, for an extended period of time, including throughout theduration of the patient's life in order to ameliorate or otherwisecontrol or limit the symptoms of the patient's disease or condition.

In certain cases wherein the patient's status does improve or does notsubstantially improve, upon the doctor's discretion the administrationof the compounds are optionally given continuously; alternatively, thedose of drug being administered is optionally temporarily reduced ortemporarily suspended for a certain length of time (i.e., a “drugholiday”). In certain embodiments, the length of the drug holiday variesbetween 2 days and 1 year, including by way of example only, 2 days, 3days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days,180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or365 days. The dose reduction during a drug holiday includes a reductionfrom about 10% to about 100%, including, by way of example only, about10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%,about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

In certain embodiments, once improvement of the patient's conditions hasoccurred, a maintenance dose is administered if necessary. In someembodiments, the dosage, e.g., of the maintenance dose, or the frequencyof administration, or both, are reduced, as a function of the symptoms,to a level at which the improved disease, disorder or condition isretained. In certain embodiments, however, patients are optionally givenintermittent treatment on a long-term basis upon any recurrence ofsymptoms.

In certain embodiments, the amount of a given agent that corresponds toan effective amount varies depending upon factors such as the particularcompound, disease or condition and its severity, the identity (e.g.,weight) of the subject or host in need of treatment. In someembodiments, the effective amount is, nevertheless, determined accordingto the particular circumstances surrounding the case, including, e.g.,the specific agent that is administered, the route of administration,the condition being treated, and the subject or host being treated. Incertain embodiments, however, doses employed for adult human treatmentis in the range of about 0.02 to about 5000 mg per day, in a specificembodiment about 1 to about 1500 mg per day. In various embodiments, thedesired dose is conveniently presented in a single dose or as divideddoses administered simultaneously (or over a short period of time) or atappropriate intervals, for example as two, three, four or more sub-dosesper day.

In some embodiments, the pharmaceutical compositions described hereinare in a unit dosage form suitable for single administration of precisedosages. In some instances, in unit dosage form, the formulation isdivided into unit doses containing appropriate quantities of one or morecompound. In certain embodiments, the unit dosage is in the form of apackage containing discrete quantities of the formulation. Non-limitingexamples are packaged tablets or capsules, and powders in vials orampoules.

In some embodiments, aqueous suspension compositions are packaged insingle-dose non-reclosable containers. In alternative embodiments,multiple-dose reclosable containers are used, in which case it istypical to include a preservative in the composition. By way of exampleonly, formulations for parenteral injection are, in some embodiments,presented in unit dosage form, which include, but are not limited toampoules, or in multi-dose containers, with an added preservative.

In certain embodiments, the daily dosages appropriate for the compoundsdescribed herein described herein are from about 0.01 to about 2.5 mg/kgper body weight. In some embodiments, an indicated daily dosage in thelarger subject, including, but not limited to, humans, is in the rangefrom about 0.5 mg to about 100 mg, conveniently administered in divideddoses, including, but not limited to, up to four times a day or inextended release form. In certain embodiments, suitable unit dosageforms for oral administration comprise from about 1 to about 50 mgactive ingredient. The foregoing ranges are merely suggestive, as thenumber of variables in regard to an individual treatment regime islarge, and considerable excursions from these recommended values are notuncommon. In certain embodiments, the dosages are altered depending on anumber of variables, not limited to the activity of the compound used,the disease or condition to be treated, the mode of administration, therequirements of the individual subject, the severity of the disease orcondition being treated, and the judgment of the practitioner.

In certain embodiments, toxicity and therapeutic efficacy of suchtherapeutic regimens are determined by standard pharmaceuticalprocedures in cell cultures or experimental animals, including, but notlimited to, the determination of the LM) (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between the toxic and therapeuticeffects is the therapeutic index and it can be expressed as the ratiobetween LD₅₀ and ED₅₀. In certain embodiments, compounds exhibiting hightherapeutic indices are preferred. In some embodiments, the dataobtained from cell culture assays and animal studies is used informulating a range of dosage for use in human. In specific embodiments,the dosage of such compounds lies within a range of circulatingconcentrations that include the ED₅₀ with minimal toxicity. In certainembodiments, the dosage varies within this range depending upon thedosage form employed and the route of administration utilized.

In certain embodiments, the disclosed compounds exhibit an increasedaffinity for a nuclear target, increased potency or increasedtherapeutic index as compared to an unmodified nuclear payload fromwhich the compound was derived. In certain embodiments, this higheraffinity, potency or therapeutic index may provide benefits, such asallowing for the administration of lower doses and thus reducedpotential for toxicity, improvement in therapeutic index and decreasedoverall cost of therapy. In certain embodiments, the daily dosagesappropriate for administration of the compounds described herein is lessthan 100% of the recommended daily dose of the unmodified nuclearpayload, or less than about 90%, or less than about 80% or less thanabout 70%, or less than about 60%, or less than about 50%, or less thanabout 40%, or from about 20% to about 90%, or from about 30% to about90%, or from about 40% to about 90%, or from about 50% to about 90%, orfrom about 60% to about 90%, or from about 70% to about 90%, or fromabout 20% to about 80%, or from about 30% to about 80%, or from about40% to about 80%, or from about 50% to about 80%, or from about 60% toabout 80%, or from about 70% to about 80%, or from about 20% to about70%, or from about 30% to about 70%, or from about 40% to about 70%, orfrom about 50% to about 70%, or from about 60% to about 70%, of therecommended daily dose of the unmodified nuclear payload.

In certain embodiments, the compounds described herein are used in thepreparation or manufacture of medicaments for the treatment of diseasesor conditions that are mediated by the enzyme poly(ADP-ribose)polymerase(PARP) or in which inhibition of the enzyme poly(ADP-ribose)polymerase(PARP) ameliorates the disease or condition. In some embodiments, amethod for treating any of the diseases or conditions described hereinin a subject in need of such treatment, involves administration ofpharmaceutical compositions containing at least one compound describedherein, or a pharmaceutically acceptable salt, pharmaceuticallyacceptable N-oxide, pharmaceutically active metabolite, pharmaceuticallyacceptable prodrug, or pharmaceutically acceptable solvate thereof, intherapeutically effective amounts to said subject.

Combination Therapy

Compounds described herein (e.g., a compound of Table 1, a compound ofFormula I or II, or any other formula disclosed herein) can also be usedin combination with other active ingredients. Such combinations areselected based on the condition to be treated, cross-reactivities ofingredients and pharmaco-properties of the combination. In oneembodiment, the disclosure provides a use of a compound as describedherein used in combination with another agent or therapy method, such asanother cancer treatment. For example, when treating cancer, thecompositions can be combined with other anti cancer compounds (such aspaclitaxel or rapamycin).

It is also possible to combine a compound of the disclosure with one ormore other active ingredients in a unitary dosage form for simultaneousor sequential administration to a patient. The combination therapy maybe administered as a simultaneous or sequential regimen. Whenadministered sequentially, the combination may be administered in two ormore administrations.

The combination therapy may provide “synergy” and “synergistic”, i.e.the effect achieved when the active ingredients used together is greaterthan the sum of the effects that results from using the compoundsseparately. A synergistic effect may be attained when the activeingredients are: (1) co-formulated and administered or deliveredsimultaneously in a combined formulation; (2) delivered by alternationor in parallel as separate formulations; or (3) by some other regimen.When delivered in alternation therapy, a synergistic effect may beattained when the compounds are administered or delivered sequentially,e.g. in separate tablets, pills or capsules, or by different injectionsin separate syringes. In general, during alternation therapy, aneffective dosage of each active ingredient is administered sequentially,i.e. serially, whereas in combination therapy, effective dosages of twoor more active ingredients are administered together. A synergisticanti-cancer effect denotes an anti-cancer effect that is greater thanthe predicted purely additive effects of the individual compounds of thecombination.

Administration of the compounds and compositions of the presentdisclosure to a patient will follow general protocols for theadministration of chemotherapeutics, taking into account the toxicity,if any. It is expected that the treatment cycles would be repeated asnecessary. It also is contemplated that various standard therapies oradjunct cancer therapies, as well as surgical intervention, may beapplied in combination with the described active agent(s). Thesetherapies include but are not limited to chemotherapy, radiotherapy,immunotherapy, gene therapy and surgery.

In some embodiments, provided herein is a method for the treatment ofcancer, comprising administering to a subject in need of treatment atherapeutically-effective amount of a compound or composition describedherein in combination with ionizing radiation or one or morechemotherapeutic agents. In some embodiments, the compound describedherein is administered simultaneously with ionizing radiation or one ormore chemotherapeutic agents. In other embodiments, the compounddescribed herein is administered sequentially with ionizing radiation orone or more chemotherapeutic agents.

In certain embodiments, provided herein is a method for the treatment ofcancer, which includes administering to a subject in need of treatment atherapeutically-effective amount of a compound or composition describedherein in combination with ionizing radiation and one or morechemotherapeutic agents. In some embodiments, the compound describedherein is administered simultaneously with ionizing radiation and one ormore chemotherapeutic agents. In other embodiments, the compounddescribed herein is administered sequentially with ionizing radiationand one or more chemotherapeutic agents.

In certain embodiments, provided herein is a method for the treatment ofcancer, which includes administering to a subject in need of treatment atherapeutically-effective amount of a compound or composition describedherein in combination with ionizing radiation. In certain embodiments,the radiation is administered at a dose of less than about 2.5 Gy perday, or about 2.0 Gy per day, or about 1.8 Gy per day, or about 1.6 Gyper day, or about 1.4 Gy per day, or about 1.2 Gy per day. In certainembodiments, a dose of less than about 2.5 Gy, or about 2.0 Gy, or about1.8 Gy, or about 1.6 Gy, or about 1.4 Gy, or about 1.2 Gy isadministered about 5 times per week. In certain embodiments, theradiation is administered at a dose of less than about 2.5 Gy per day,or about 2.0 Gy per day, or about 1.8 Gy per day, or about 1.6 Gy perday, or about 1.4 Gy per day, or about 1.2 Gy per day. In certainembodiments, a dose of less than about 2.5 Gy, or about 2.0 Gy, or about1.8 Gy, or about 1.6 Gy, or about 1.4 Gy, or about 1.2 Gy isadministered about 6 times per week. It is contemplated that byadministering radiation in combination with a compound or compositiondescribed herein, prostate specific chemical prostatectomy can beachieved while avoiding detrimental side effects, such as the impotenceand incontinence of surgical prostatectomy due to disruption of vesselsand nerves.

Cancer therapies can also include a variety of combination therapieswith both chemical and radiation based treatments. Combinationchemotherapies include the use of chemotherapeutic agents such as,cisplatin, etoposide, irinotecan, camptostar, topotecan, paclitaxel,docetaxel, epothilones, taxotere, tamoxifen, 5-fluorouracil,methoxtrexate, temozolomide, cyclophosphamide, SCH 66336, R115777,L778,123, BMS 214662, IRESSA® (gefitinib), TARCEVAR® (erlotinibhydrochloride), antibodies to EGFR, GLEEVEC® (imatinib), intron, ara-C,adriamycin, cytoxan, gemcitabine, uracil mustard, chlormethine,ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine,triethylenethiophosphoramine, busulfan, carmustine, lomustine,streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine,6-thioguanine, fludarabine phosphate, pentostatine, vinblastine,vincristine, vindesine, bleomycin, doxorubicin, dactinomycin,daunorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin,Mitomycin-C, L-Asparaginase, teniposide, 17α-Ethinylestradiol,Diethylstilbestrol, testosterone, prednisone, fluoxymesterone,dromostanolone propionate, testolactone, megestrolacetate,methylprednisolone, methyltestosterone, prednisolone, triamcinolone,chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine,medroxyprogesterone acetate, leuprolide, flutamide, toremifene,goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane,mitoxantrone, levamisole, navelbene, anastrazole, letrazole,capecitabine, reloxafine, droloxafine, hexamethylmelamine, Avastin,herceptin, Bexxar, Velcade, Zevalin, Trisenox, Xeloda, Vinorelbine,Porfimer, Erbitux® (cetuximab), Liposomal, Thiotepa, Altretamine,Melphalan, Trastuzumab, Lerozole, Fulvestrant, Exemestane, Ifosfomide,Rituximab, C225, Campath, carboplatin, procarbazine, mechlorethamine,cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil,busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin,bleomycin, plicomycin, mitomycin, etoposide (VP 16), tamoxifen,raloxifene, estrogen receptor binding agents, paclitaxel, gemcitabine,navelbine, farnesyl-protein transferase inhibitors, transplatinum,5-fluorouracil, vincristine, vinblastine and methotrexate, or any analogor derivative variant of the foregoing.

Other factors that cause DNA damage, such as radiotherapy, have beenused extensively include what are commonly known as gamma-rays, X-rays,and/or the directed delivery of radioisotopes to tumor cells. Otherforms of DNA damaging factors are also contemplated such as microwavesand UV-irradiation. It is most likely that all of these factors affect abroad range of damage on DNA, on the precursors of DNA, on thereplication and repair of DNA, and on the assembly and maintenance ofchromosomes. Dosage ranges for X-rays range from daily doses of 50 to200 roentgens for prolonged periods of time (e.g., 3 to 4 weeks), tosingle doses of 2000 to 6000 roentgens. Dosage ranges for radioisotopesvary widely, and depend on the half-life of the isotope, the strengthand type of radiation emitted, and the uptake by the neoplastic cells.The terms “contacted” and “exposed,” when applied to a cell, are usedherein to describe the process by which a therapeutic construct and achemotherapeutic or radiotherapeutic agent are delivered to a targetcell or are placed in direct juxtaposition with the target cell. Toachieve cell killing or stasis, both agents are delivered to a cell in acombined amount effective to kill the cell or prevent it from dividing.

Immunotherapeutics, generally, rely on the use of immune effector cellsand molecules to target and destroy cancer cells. The immune effectormay be, for example, an antibody specific for some marker on the surfaceof a tumor cell. The antibody alone may serve as an effector of therapyor it may recruit other cells to actually affect cell killing. Theantibody also may be conjugated to a drug or toxin (chemotherapeutic,radionucleotide, ricin A chain, cholera toxin, pertussis toxin, etc.)and serve merely as a targeting agent. Alternatively, the effector maybe a lymphocyte carrying a surface molecule that interacts, eitherdirectly or indirectly, with a tumor cell target. Various effector cellsinclude cytotoxic T cells and NK cells.

Immunotherapy, thus, could be used as part of a combined therapy, inconjunction with gene therapy. The general approach for combined therapyis discussed below. Generally, the tumor cell must bear some marker thatis amenable to targeting, i.e., is not present on the majority of othercells. Many tumor markers exist and any of these may be suitable fortargeting in the context of the present disclosure. Common tumor markersinclude carcinoembryonic antigen, prostate specific antigen, urinarytumor associated antigen, fetal antigen, tyrosinase (p97), gp68, TAG-72,HMFG, Sialyl Lewis Antigen, MucA, MucB, PLAP, estrogen receptor, lamininreceptor, erb B and p155.

In yet another embodiment, the secondary treatment is a secondary genetherapy in which a therapeutic polynucleotide is administered before,after, or at the same time a first chemotherapeutic agent. Delivery ofthe chemotherapeutic agent in conjunction with a vector encoding a geneproduct will have a combined anti-hyperproliferative effect on targettissues.

Approximately 60% of persons with cancer will undergo surgery of sometype, which includes preventative, diagnostic or staging, curative andpalliative surgery. Curative surgery is a cancer treatment that may beused in conjunction with other therapies, such as the treatment of thepresent disclosure, chemotherapy, radiotherapy, hormonal therapy, genetherapy, immunotherapy and/or alternative therapies. Curative surgeryincludes resection in which all or part of cancerous tissue isphysically removed, excised, and/or destroyed. Tumor resection refers tophysical removal of at least part of a tumor. In addition to tumorresection, treatment by surgery includes laser surgery, cryosurgery,electrosurgery, and microscopically controlled surgery (Mohs' surgery).It is further contemplated that the present disclosure may be used inconjunction with removal of superficial cancers, precancers, orincidental amounts of normal tissue.

Administration of the compound or composition as described herein mayprecede or follow the other anti-cancer agent or treatment by intervalsranging from minutes to weeks. In embodiments where the otheranti-cancer agent and expression construct are applied separately, onewould generally ensure that a significant period of time did not elapsebetween the time of each delivery, such that the agent and expressionconstruct would still be able to exert an advantageously combined effecton a cell. For example, in such instances, it is contemplated that onemay contact a cell, tissue or organism with two, three, four or moremodalities substantially simultaneously (i.e., within less than about aminute) with the active agent(s). In other aspects, one or more agentsmay be administered within about 1 minute, about 5 minutes, about 10minutes, about 20 minutes about 30 minutes, about 45 minutes, about 60minutes, about 2 hours, about 3 hours, about 4 hours, about 6 hours,about 8 hours, about 9 hours, about 12 hours, about 15 hours, about 18hours, about 21 hours, about 24 hours, about 28 hours, about 31 hours,about 35 hours, about 38 hours, about 42 hours, about 45 hours, to about48 hours or more prior to and/or after administering the activeagent(s). In certain other embodiments, an agent may be administeredwithin from about 1 day, about 2 days, about 3 days, about 4 days, about5 days, about 6 days, about 8 days, about 9 days, about 12 days, about15 days, about 16 days, about 18 days, about 20 days, to about 21 daysprior to and/or after administering the active agent(s). In somesituations, it may be desirable to extend the time period for treatmentsignificantly, however, where several weeks (e.g., about 1, about 2,about 3, about 4, about 6, or about 8 weeks or more) lapse between therespective administrations.

Kits

Kits for use to achieve anti-cancer effects comprising a compound orcomposition described herein are provided. In certain embodiments, thekit comprises a unit dose of a compound or composition described hereinand instructions for administering the same. In certain aspects, the kitfurther comprises a second drug suitable for anti-cancer therapy, orinstructions for co-administering an additional anti cancer therapy(such as radiation or gene therapy). In another aspect, kits for use toachieve anti-cancer effects comprise a low dose (e.g., less than about500 mg/day, or less than about 400 mg/day, or less than about 300mg/day, or less than about 200 mg/day) of a compound or compositiondescribed herein and a second drug suitable for anti-cancer therapy. Inyet another variation, kits for use to achieve anti-cancer effectscomprise a high dose (e.g., greater than about 500 mg/day) of a compoundor composition as described herein and a second drug suitable foranti-cancer therapy.

Methods of Manufacturing a Medicament

In a further aspect of the disclosure, use of the compounds andcompositions described herein in the manufacture of a medicament isprovided. In particular, the manufacture of a medicament for use in thetreatment of cancer, or diseases or conditions which can be mediated, atleast in part, by blocking DNA repair and/or transcription activation,such as by inhibition of PARP, are provided. Further, pharmaceuticalcompositions of a compound described herein are also intended for use inthe manufacture of a medicament for use in treatment of diseases orconditions which can be mediated, at least in part, by inhibition ofPARP.

EXAMPLES

The disclosure is further illustrated by the following examples. Theexamples below are non-limiting are merely representative of variousaspects of the disclosure. Solid and dotted wedges within the structuresherein disclosed illustrate relative stereochemistry, with absolutestereochemistry depicted only when specifically stated or delineated.

Compounds as disclosed herein, or any formula or sub-formula describedherein, can be synthesized using standard synthetic techniques known tothose of skill in the art. Compounds of the present disclosure can besynthesized using the general synthetic procedures set forth in GeneralMethods 1-5 Synthetic Examples.

Where it is desired to obtain a particular enantiomer of a compound,this may be accomplished from a corresponding mixture of enantiomersusing any suitable conventional procedure for separating or resolvingenantiomers. Thus, for example, diastereomeric derivatives may beproduced by reaction of a mixture of enantiomers, e.g., a racemate, andan appropriate chiral compound. The diastereomers may then be separatedby any convenient means, for example by crystallization and the desiredenantiomer recovered. In another resolution process, a racemate may beseparated using chiral High Performance Liquid Chromatography.Alternatively, if desired a particular enantiomer may be obtained byusing an appropriate chiral intermediate in one of the processesdescribed.

Chromatography, recrystallization and other conventional separationprocedures may also be used with intermediates or final products whereit is desired to obtain a particular isomer of a compound or tootherwise purify a product of a reaction.

General Information

¹H NMR spectra and ¹³C NMR spectra were recorded on Varian 400 MHz orBroker Avance III 500 MHz spectrometers. Spectra are referenced toresidual chloroform (δ 7.26, ¹H), DMSO (δ 2.54, ¹H) or methanol (δ 3.34,¹H) unless otherwise noted. Chemical shifts are reported in ppm (δ);multiplicities are indicated by s (singlet), d (doublet), t (triplet), q(quartet), quint (quintet), sext (sextet), m (multiplet) and br (broad).Coupling constants, J, are reported in Hertz. Silica gel chromatographywas performed using a Teledyne Isco CombiFlash® Rf+ instrument usingHi-Purit Silica Flash Cartridges (National Chromatography Inco) orRediSep Rf Gold C18 Cartridges (Teledyne Isco). Analytical HPLC wasperformed on a Waters ACQUITY UPLC with a photodiode array detectorusing and a Waters ACQUITY BEH Shield RPC18 (2.1×50 mm, 1.7 μm) column.Analytical LCMS was performed on a Waters ACQUITY UPLC with a Waters3100 mass detector. Chiral HPLC was performed on a Waters Alliance e2695with a photodiode array detector using Daicel Chiralpak® AD-H,Chiralpak® IA, Chiralpak® IB, Chiralpak® IC, Chiralcel® OD-H orChiralcel® OJ-H columns. Optical rotations were obtained on a JascoP-2000 digital polarimeter and are reported as [α]_(D) ^(T) temperature(T), concentration (c=g/100 mL) and solvent. Commercially availablereagents and solvents were used as received unless otherwise indicated.

General Methods General Method 1

Olaparib-containing analogs can be prepared following the methoddescribed by Menear et al. (Menear, K. A. et al. J. Med. Chem. 2008, 51,6581-6591).

General Method 2

Rucaparib-containing analogs can be prepared following the methoddescribed by Gillmore et. al. (Gillmore, A. T. el al. Org. Process Res.Dev. 2012, 16, 1897-1904).

General Method 3

Talazoparib-containing analogues can be prepared following the methoddescribed by Wang et al. (Wang, B. et al. J. Med. Chem. 2016, 59,335-357).

General Method 4

DHT-containing intermediates can be prepared by treating commerciallyavailable DHT (XX) with the desired tethering group to affordintermediates such as XXI. When R=CO₂Me, the ester group can be mildlysaponified to provide the carboxylate intermediates A. When R=OAc, mildsaponification releases the primary alcohol which can then be treatedwith methanesulfonyl chloride to afford intermediates B.

General Method 5

Enzalutamide-containing intermediates can be prepared starting withcarboxylic acid XXII (Jadhavar, et al. Bioorg. Med. Chem. Lett. 2016,26, 5222-5228) and coupling with the desired tethering group to affordintermediates such as XXIII. When R=CO₂Me, the ester group can be mildlysaponified to provide the carboxylate intermediates C. When R=OAc, mildsaponification releases the primary alcohol which can then be treatedwith methanesulfonyl chloride to afford intermediates D.

General Method 6

CC-115 containing analogs can be prepared following the method describedby Mortensen et al. (Mortensen, D. S. et al. J. Med. Chem. 2015, 58,5599-5608).

Synthetic Examples Example S-1. Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexyl)benzamide(Compound 1.1)

Step-1: Preparation of(Z)-2-fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)benzonitrile

To a stirred solution of dimethyl 3-oxo-1,3-dihydroisobenzofuran-1-ylphosphonate (10 g, 41.3 mmol) and 2-fluoro-5-formylbenzonitrile (6.15 g,41.3 mmol) in THE (50 mL) was added triethylamine (5.76 mL, 41.3 mmol)at 0° C. slowly. The resultant mixture was stirred at RT for 16 h. Thereaction was monitored by TLC. Upon completion, water (200 mL) was addedand the resulting precipitate was filtered via a Buchner funnel. Theproduct obtained was washed with water (50 mL), hexanes (50 mL), diethylether (30 mL), dried under vacuum afford to afford(Z)-2-fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)benzonitrile(50:50 mixture of E and Z isomers) which was used in next step withoutfurther purification. LC-MS: 266 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.13(1H, m), 8.05 (1H, m), 7.98 (1H, m), 7.79 (2H, m), 7.61 (1H, m), 7.30(1H, m), 6.35 (1H, s).

Step-2: Preparation of 2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoic acid

To a stirred suspension of(Z)-2-fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)benzonitrile(3.7 g, 13.9 mmol) in water (20 mL) was added 13N aqueous NaOH (5 mL)and the mixture was heated under nitrogen at 90° C. for 16 h. Thereaction mixture was then cooled to 70° C. and hydrazine hydrate (10 mL)was added and stirred for 16 hours at 70° C. The reaction was monitoredby TLC. Upon completion, the reaction was cooled to RT and acidifiedusing 2N aqueous HCl (pH 1-2) at 0-5° C. to obtain a precipitate. Theslurry was filtered via a Buchner funnel, washed with water (50 mL),hexanes (50 mL), diethyl ether (30 mL) and dried under vacuum afford toafford 2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoicacid. LC-MS 299 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 13.22 (1H, br. s),12.61 (1H, s), 8.27 (1H, m), 7.99-7.81 (4H, m), 7.59 (1H, m), 7.25 (1H,m), 4.36 (2H, s).

Step-3: Preparation of tert-butyl4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazine-1-carboxylate

To a stirred suspension of2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoic acid (0.1g, 0.335 mmol) in DMA (4 mL) was added HBTU (0.15 g, 0.402 mmol, 1.2 eq)at 0° C. and the mixture was stirred for 10 min. DIPEA (0.17 mL, 1 mmol,3 eq) and tert-butyl piperazine-1-carboxylate (0.075 g, 0.402 mmol, 1.2eq) were then successively added to the reaction mixture at 0° C. andthe resultant reaction mixture was stirred at RT for 75 min. Thereaction was monitored by TLC. After completion, water (10 mL) was addedand the resulting precipitate was filtered via a Buchner funnel. Theproduct obtained was washed with water (10 mL×2) and n-pentane (10mL×2), dried under reduced pressure to afford tert-butyl4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazine-1-carboxylatewhich was taken to next step without further purification. LC-MS 467[M+H]⁺.

Step-4: Preparation of4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of tert-butyl4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazine-1-carboxylate(0.150 g, 0.321 mmol) was added 2N aqueous HCl in MeOH (5 mL) at RT andthe mixture was stirred at RT for 16 h. The reaction was monitored byLC-MS. Upon completion, the reaction mixture was concentrated underreduced pressure to afford4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one as anHCl salt. LC-MS 367 [M+H]+.

Step-1a: Preparation of methyl 6-aminohexanoate

To 6-aminohexanoic acid (10.0 g, 76.23 mmol) was added 2N aqueous HCl inMeOH (30 mL) at RT and the mixture was stirred at RT for 16 h. Thereaction was monitored by ¹H NMR. Upon completion, the reaction mixturewas concentrated under reduced pressure to afford methyl6-aminohexanoate which was triturated with diethyl ether and pentane(1:3) to obtain the title compound as a hydrochloride salt. ¹H NMR (400MHz, MeOD-d₄) δ 3.66 (s, 3H), 2.92 (t, J=7.67 Hz, 2H), 2.37 (t, J=7.45Hz, 2H), 1.62-1.71 (m, 6H), 1.37-1.45 (m, 2H).

Step-1b: Preparation of methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-2-fluorobenzamido)hexanoate

To a stirred solution of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoicacid (3 g, 6.64 mmol) in DMF (25 mL) was added HATU (5.05 g, 13.29 mmol,2 eq) at 0° C. and the mixture was stirred for 30 min. DIPEA (5.78 mL,33.2 mmol, 5 eq) and 6-aminohexanoate (1.81 g, 9.96 mmol, 1.5 eq) werethen successively added to the reaction mixture and the mixture wasstirred at RT for 4 h. The reaction was monitored by TLC and LC-MS.After completion, the reaction was diluted with EtOAc (250 mL). Theorganic layer was washed with saturated aqueous NaHCO₃ (100 mL),saturated aqueous NH₄Cl (100 mL), water (100 mL), brine (50 mL), driedover anhydrous Na₂SO₄, filtered and concentrated under reduced. Thecrude material was purified by CombiFlash to afford methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoate.LC-MS 579 [M+H]⁺.

Step-1c: Preparation of6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-2-fluorobenzamido)hexanoic acid

To a stirred solution of methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoate(3.5 g, 6.05 mmol) in THE (30 mL) and MeOH (15 mL) was added LiOH.10H₂O(2.53 g, 60.5 mmol, 10 eq). The solids were dissolved in water (5 mL) atRT and the mixture was stirred at RT for 16 h. The reaction wasmonitored by TLC. After completion, the reaction mixture wasconcentrated under reduced pressure. The residue obtained was dilutedwith water (30 mL) and acidified using 2N aqueous HCl (pH ˜2). Theaqueous layer was then extracted with EtOAc (300 mL×3). The combinedorganic layers were washed with brine (200 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure to afford6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoicacid which was taken to next step without further purification. LC-MS565 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.51 (br. s., 1H), 8.41 (d,J=7.83 Hz, 1H), 8.30 (s, 1H), 8.08 (d, J=8.31 Hz, 1H), 7.75 (t, J=7.83Hz, 1H), 7.42 (d, J=10.27 Hz, 1H), 7.33 (d, J=7.83 Hz, 1H), 3.25 (d,J=6.36 Hz, 2H), 2.72-66 (m, 4H), 2.51-2.4 (m, 2H), 2.21 (t, J=7.58 Hz,2H), 1.54 (s, 6H), 1.34 (d, J=6.85 Hz, 2H).

Step-5: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1yl)-6-oxohexyl)benzamide (Compound 1.1)

To a stirred solution of6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoicacid (0.150 g, 0.265 mmol) in DMA (4 mL) was added HBTU (0.12 g, 0.319mmol, 1.2 eq) at 0° C. and the mixture was stirred for 10 min. DIPEA(0.16 mL, 0.93 mmol, 3.5 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride salt (0.128 g, 0.319 mmol, 1.2 eq) were then successivelyadded to the reaction mixture and the mixture was stirred at RT for 1.5h. The reaction was monitored by TLC and LC-MS. After completion, water(10 mL) was added and the resulting precipitate was filtered via aBuchner funnel. The product obtained was washed with water (10 mL×2) andn-pentane (10 mL×2), dried under reduced pressure to obtain a crudeproduct which was purified by reversed-phase HPLC to afford Compound1.1. LC-MS 913 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD-d₄) δ 8.36 (d, J=7.8 Hz,1H), 8.16 (d, J=7.8 Hz, 2H), 7.99 (dd, J=7.9, 2.1 Hz, 1H), 7.94 (dd,J=8.3, 3.4 Hz, 1H), 7.91-7.78 (m, 3H), 7.47 (p, J=4.5, 4.0 Hz, 1H), 7.36(td, J=9.2, 8.8, 5.1 Hz, 3H), 7.15 (td, J=9.0, 2.4 Hz, 1H), 4.37 (s,2H), 3.83-3.69 (m, 2H), 3.69-3.62 (m, 2H), 3.50 (dt, J=9.2, 4.9 Hz, 2H),3.42 (q, J=6.4, 6.0 Hz, 2H), 3.35 (m, 2H), 3.28 (m, 2H), 2.48 (t, J=7.3Hz, 1H), 2.41 (t, J=7.5 Hz, 1H), 1.67 (q, J=7.5 Hz, 4H), 1.59 (s, 6H),1.53-1.40 (m, 2H).

Example S-2. Preparation of4-(3-(4-(6-(((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one(Compound 1.2)

Step-1: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol

To a stirred solution of (5 S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(3 g, 10.3 mmol) in benzene (100 mL) was added PTSA (0.982 g, 5.1 mmol,0.5 eq) followed by addition of ethylene glycol (2.88 g, 51.61 mmol, 5eq) at RT. The resultant mixture was heated to 120° C. using aDean-Stark apparatus for 16 h. The reaction was monitored by TLC. Uponcompletion, the reaction mixture was diluted with water (200 mL) andextracted with EtOAc (350 mL). The organic layer was washed withsaturated NaHCO₃ solution (100 mL), water (200 mL), brine (100 mL) driedover Na₂SO₄, filtered and concentrated under reduced pressure to afford(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol.LC-MS 335 [M+H]+. ¹H NMR (400 MHz, DMSO-d₆) δ 4.40 (d, J=5.26 Hz, 1H),3.81 (s, 3H), 3.37-3.45 (m, 2H), 1.82 (br. s., 2H), 1.70 (d, J=12.72 Hz,2H), 1.59 (d, J=13.15 Hz, 2H), 1.52 (d, J=10.52 Hz, 3H), 1.35 (br. s.,1H), 1.23-1.33 (m, 3H), 1.03-1.21 (m, 5H), 0.87 (dt, J=7.89, 19.95 Hz,4H), 0.76 (s, 3H), 0.61 (s, 3H).

Step-2a: Preparation of 2-(6-chlorohexyloxy)tetrahydro-21-1 pyran

To a stirred solution of 6-chlorohexan-1-ol (10 g, 73.158 mmol) indiethyl ether (100 mL) was added pTSA (0.05 g, 0.365 mmol, 0.005 eq)followed by addition of 3,4-dihydro-2H-pyran (8.4 g, 102.48 mmol, 1.4eq). The resulting reaction mixture was stirred at RT for 16 h. Thereaction was monitored by TLC. Upon completion, the reaction mixture wasdiluted with diethyl ether (350 mL). The organic layer was washed with20% KOH solution (100 mL), water (300 mL), brine (200 mL) dried overNa₂SO₄, filtered and concentrated under reduced pressure to afford2-(6-chlorohexyloxy)tetrahydro-2H-pyran. ¹H NMR (400 MHz, DMSO-d₆) δ4.50 (br. s., 1H), 4.25 (t, J=6.80 Hz, 2H), 4.20 (s, 1H), 4.00-4.08 (m,2H), 3.66-3.74 (m, 2H), 3.59 (dd, J=6.36, 16.01 Hz, 2H), 2.66 (s, 2H),1.81-1.90 (m, 2H), 1.67 (br. s., 1H), 1.38-1.45 (m, 2H), 1.20-1.27 (m,2H), 1.17 (t, J=7.02 Hz, 2H).

Step-2: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(6-(tetrahydro-2H-pyran-2-yloxy)hexyloxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane]

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol(2 g, 5.97 mmol) in xylene (50 mL) was added NaNH₂ (50% suspension intoluene, 1.4 mL) and the mixture was heated at 150° C. for 1 h. Thereaction mixture was gradually cooled to RT, 2-(6-chlorohexyloxy)tetrahydro-2H-pyran (2.8 mL) was added to it and the resultant mixturewas again heated to 150° C. for 16 h. The reaction was monitored by TLC.

Upon completion, the mixture was cooled to RT, quenched slowly with icecold water (250 mL) and extracted with EtOAc (300 mL). The organic layerwas washed with water (100 mL×2), brine (100 mL) dried over Na₂SO₄,filtered and concentrated under reduced pressure to obtain a crudeproduct which was purified by CombiFlash chromatography (Teledyne IscoRf+); compound eluting 30% EtOAc/hexane to afford(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(6-(tetrahydro-2H-pyran-2-yloxy)hexyloxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane].¹H NMR (400 MHz, CDCl3) δ 4.57 (d, J=3.95 Hz, 1H), 3.91-3.95 (m, 4H),3.82-3.91 (m, 2H), 3.73 (td, J=6.80, 9.65 Hz, 2H), 3.42-3.53 (m, 3H),3.33-3.42 (m, 3H), 3.26 (t, J=8.55 Hz, 2H), 1.91-2.01 (m, 2H), 1.75-1.90(m, 3H), 1.46-1.74 (m, 13H), 1.31-1.42 (m, 6H), 1.07-1.28 (m, 4H),0.83-0.97 (m, 4H), 0.81 (s, 4H), 0.70-0.76 (m, 4H), 0.07 (s, 3H), 0.00(s, 3H).

Step-3: Preparation of(5S,8R,9S,10S,13S,14S,17S)-17-(6-hydroxyhexyloxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(6-(tetrahydro-2H-pyran-2-yloxy)hexyloxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane](1 g, 1.92 mmol) in THE (25 mL) water (5 mL) was added 6N HCl (15 mL) atRT and the resultant reaction mixture was stirred at RT for 16 h. Thereaction was monitored by TLC. Upon completion, the reaction mixture wasdiluted with water (150 mL) and basified using with saturated NaHCO₃solution (pH ˜8). The aqueous layer was then extracted with EtOAc (200mL×3). The organic layer was washed with saturated NaHCO₃ solution (100mL), water (100 mL), brine (100 mL) dried over Na₂SO₄, filtered andconcentrated under reduced pressure to afford(5S,8R,9S,10S,13S,14S,17S)-17-(6-hydroxyhexyloxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-onewhich was used in the next step without further purification. ¹H NMR(400 MHz, CDCl3) δ 4.90 (br. s., 1H), 4.12 (d, J=7.45 Hz, 1H), 4.00 (br.s., 1H), 3.64 (t, J=6.58 Hz, 3H), 3.49-3.58 (m, 1H), 3.37-3.47 (m, 3H),3.27 (t, J=8.55 Hz, 1H), 2.22-2.43 (m, 3H), 2.10 (br. s., 1H), 1.93-2.07(m, 2H), 1.90 (d, J=11.84 Hz, 2H), 1.82 (d, J=7.89 Hz, 2H), 1.70 (d,J=10.09 Hz, 2H), 1.48 (d, J=4.38 Hz, 2H), 1.30-1.42 (m, 8H), 1.22-1.29(m, 2H), 1.01 (s, 3H), 0.77 (s, 3H).

Step-4: Preparation of6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-(6-hydroxyhexyloxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(0.1 g, 0.256 mmol) in acetone (5 mL) was added Jones reagent (0.5 mL)at 0° C. dropwise over a period of 20 min. The resultant mixture wasstirred at 0° C. for 5 min. The reaction was monitored by TLC. Aftercompletion, water (10 mL) was added and the resulting precipitate wasfiltered over Buchner funnel. The product obtained was washed with water(5 mL×2) and n-pentane (5 mL×2), dried under vacuum to afford 6((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid which was taken to next step without further purification. ¹H NMR(400 MHz, DMSO-d₆) δ 3.19-3.28 (m, 3H), 2.24-2.35 (m, 2H), 2.18 (t,J=7.24 Hz, 2H), 2.08 (d, J=13.59 Hz, 2H), 1.84-1.95 (m, 3H), 1.79 (d,J=10.52 Hz, 2H), 1.61 (d, J=13.59 Hz, 2H), 1.40-1.54 (m, 8H), 1.22-1.38(m, 8H), 1.19 (br. s., 1H), 1.09-1.18 (m, 2H), 0.97 (s, 3H), 0.69 (s,3H).

Step-5: Preparation of4-(3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one(Compound 2)

To a stirred solution of6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid (0.1 g, 0.247 mmol) in DMA (5 mL) was added HBTU (0.112 g, 0.296mmol, 1.2 eq) at 0° C. and the resulting reaction mixture was stirredfor 10 min. DIPEA (0.94 mL, 0.544 mmol, 2.2 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one (0.108g, 0.296 mmol, 1.2 eq) were then successively added to the mixture andthe mixture was stirred at RT for 1 h. The reaction was monitored by TLC& LC-MS. After completion, water (10 mL) was added and the resultingprecipitate was filtered via a Buchner funnel. The product obtained waswashed with water (5 mL×2) and n-pentane (5 mL×2), dried under vacuum toobtain a crude product which was purified by reverse phase HPLC toafford Compound 1.2. LC-MS 753 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.37(d, J=7.6 Hz, 1H), 7.99-7.91 (m, 1H), 7.91-7.79 (m, 2H), 7.48 (t, J=6.4Hz, 1H), 7.36 (d, J=6.1 Hz, 1H), 7.21-7.11 (m, 1H), 4.38 (s, 2H),3.82-3.61 (m, 4H), 3.47 (ddt, J=22.1, 14.2, 6.3 Hz, 4H), 3.37-3.33 (m,2H), 2.53-2.30 (m, 4H), 2.26-2.16 (m, 1H), 2.08-1.95 (m, 3H), 1.88 (t,J=11.6 Hz, 1H), 1.75-1.50 (m, 10H), 1.50-1.13 (m, 12H), 1.05 (s, 3H),1.01-0.86 (m, 2H), 0.76 (d, J=9.0 Hz, 4H).

Example S-3. Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(6-((4-((8S,9R)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-3-oxo-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-8-yl)phenyl)amino)-6-oxohexyl)benzamide(Compound 1.3)

Step-1a: Preparation of methyl5-fluoro-2-(2-(1-methyl-1H-1,2,4-triazol-5 yl)acetyl)-3-nitrobenzoate

(Z)-6-fluoro-3-((1-methyl-1H-1,2,4-triazol-5-yl)methylene)-4-nitroisobenzofuran-1(3H)-one(10 g, 34.48 mmol) was added to a reaction vessel containing 2N aqueousHCl in MeOH (50 mL) at RT and the mixture was stirred at RT for 16 h.The reaction was monitored by TLC. Upon completion, the reaction mixturewas concentrated under reduced pressure to obtain a sticky solid whichwas further lyophilized to afford methyl5-fluoro-2-(2-(1-methyl-1H-1,2,4-triazol-5-yl) acetyl)-3-nitrobenzoateas a hydrochloride salt. LC-MS 323 [M+H]⁺.

Step-1b′: Preparation of 4-aminobenzaldehyde

To a stirred solution of 4-nitrobenzaldehyde (5 g, 33.1 mmol) in ethanol(50 mL) was added SnCl₂ (37.35 g, 165.5 mmol, 5 eq) at 0° C. and themixture was heated at 80° C. for 1 h. The reaction was monitored by TLC.Upon completion, the reaction mixture was concentrated under reducedpressure to afford a crude residue which was suspended in water (100 mL)and basified using NaHCO₃ solution (pH˜8). The aqueous layer was thenextracted with EtOAc (500 mL×3). The combined organic layers were washedwith saturated NaHCO₃ solution (300 mL), water (200 mL), brine (150 mL)dried over Na₂SO₄, filtered and concentrated under reduced pressure toafford 4-aminobenzaldehyde which was used in the next step withoutfurther purification. LC-MS 122 [M+H]⁺.

Step-1b: Preparation of methyl2-(4-aminophenyl)-7-fluoro-3-(1-methyl-1H-1,2,4-triazol-5-yl)-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate

To a stirred solution of methyl5-fluoro-2-(2-(1-methyl-1H-1,2,4-triazol-5-yl) acetyl)-3-nitrobenzoatehydrochloride salt (5.6 g, 15.6 mmol) in THE (60 mL) MeOH (10 mL) wasadded 4-aminobenzaldehyde (3.8 g, 31.2 mmol, 2 eq). Titanium (III)chloride (20% w/v solution in 2N-HCl (50 mL) was then added to themixture at RT dropwise over a period of 20 min and the mixture wasstirred at 50° C. for 3 h. The reaction was monitored by TLC. Uponcompletion, the solvent is removed under reduced pressure to obtaincrude residue which was dissolved in water (300 mL) and basified usingsaturated NaHCO₃ solution (pH˜8). The aqueous layer was then extractedwith EtOAc (400 mL×3). The combined organic layers were washed saturatedNaHCO₃ solution (300 mL), water (300 mL), brine (100 mL) dried overNa₂SO₄, filtered and concentrated under reduced pressure to afford acrude product which was purified by CombiFlash chromatography (TeledyneIsco Rf+); compound eluting 40% EtOAc/hexane to afford methyl2-(4-aminophenyl)-7-fluoro-3-(1-methyl-1H-1,2,4-triazol-5-yl)-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate.LC-MS 396 [M+H]⁺.

Step-1c: Preparation of8-(4-aminophenyl)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]phthalazin-3(7H)-one

To a stirred suspension of methyl2-(4-aminophenyl)-7-fluoro-3-(1-methyl-1H-1,2,4-triazol-5-yl)-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate(1.5 g, 37.9 mmol) in methanol (20 mL) was added hydrazine hydrate (3mL) at 0° C. and the resultant mixture was stirred at RT for 4 h. Thereaction was monitored by TLC. After completion, the reaction mixturewas concentrated under reduced pressure and water (20 mL) was added. Theresulting precipitate was filtered over Buchner funnel. The productobtained was washed with water (10 mL×2) and n-pentane (10 mL×2), driedunder reduced pressure to afford a crude product which was purified byreverse phase chromatography to afford of8-(4-aminophenyl)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]phthalazin-3(7H)-oneas a racemic mixture. The compound obtained was further separated bychiral chromatography to afford4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(6-(4-((8S,9R)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-3-oxo-3,7,8,9-tetrahydro-2H-pyrido[4,3,2-de]phthalazin-8-yl)phenylamino)-6-oxohexyl)benzamide(PK-1) (0.300 g) and4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(6-(4-((8S,9R)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-3-oxo-3,7,8,9-tetrahydro-2H-pyrido[4,3,2-de]phthalazin-8-yl)phenylamino)-6-oxohexyl)benzamide(PK-2).

(PK-1 was used for Compound 1.3 and PK-2 was used for ent-Compound 1.3)

PK-1: LC-MS 378 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 12.30 (s, 1H), 7.78(s, 1H), 7.56 (s, 1H), 7.02 (t, J=6.7 Hz, 3H), 6.90 (d, J=11.2 Hz, 1H),6.45 (d, J=7.9 Hz, 2H), 5.10 (s, 2H), 4.85 (d, J=11.4 Hz, 1H), 4.72 (d,J=11.4 Hz, 1H), 3.62 (s, 3H).

PK-2: LC-MS 378 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 12.29 (s, 1H), 7.78(s, 1H), 7.56 (s, 1H), 7.02 (t, J=6.7 Hz, 3H), 6.89 (d, J=11.2 Hz, 1H),6.44 (d, J=7.9 Hz, 2H), 5.10 (s, 2H), 4.85 (d, J=11.4 Hz, 1H), 4.71 (d,J=11.4 Hz, 1H), 3.61 (s, 3H).

Step-1: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-S,S-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-2-fluoro-N-(6-(4-(5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5yl)-3-oxo-3,7,8,9-tetrahydro-2H pyrido[4,3,2-de]phthalazin-8yl)phenylamino)-6-oxohexyl)benzamide (Compound 1.3)

To a stirred solution of6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoicacid (0.125 g, 0.221 mmol) in DMF (7 mL) was added HATU (0.168 g, 0.442mmol, 2 eq) at 0° C. and the resulting mixture was stirred for 30 min.DIPEA (0.19 mL, 1.11 mmol, 5 eq) and8-(4-aminophenyl)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-dc]phthalazin-3(7H)-one(PK-1) (0.10 g, 0.265 mmol, 1.2 eq) were then successively added to thereaction mixture and the resultant mixture was stirred at RT for 4 h.The reaction was monitored by TLC. Upon completion, the reaction mixturewas diluted with water (50 mL) and extracted using EtOAc (100 mL×2). Thecombined organic layer were washed with saturated NaHCO₃ solution (100mL), saturated NH₄Cl solution (100 mL), water (100 mL), brine (50 mL)dried over Na₂SO₄, filtered and concentrated under reduced pressure toafford a crude product which was purified by reverse phase HPLC toafford Compound 1.3. LC-MS 924 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD-d₄) δ 8.15(d, J=9.1 Hz, 2H), 7.97 (d, J=8.5 Hz, 1H), 7.87 (d, J=3.0 Hz, 1H), 7.82(t, J=8.8 Hz, 1H), 7.56-7.48 (m, 2H), 7.34 (d, J=8.4 Hz, 4H), 7.19 (dd,J=7.1, 4.7 Hz, 1H), 6.90 (d, J=10.4 Hz, 1H), 3.58 (d, J=3.0 Hz, 3H),3.46-3.36 (m, 2H), 2.38 (d, J=7.6 Hz, 2H), 1.93 (s, 2H), 1.80-1.64 (m,6H), 1.59 (d, J=2.8 Hz, 6H), 1.48 (ddt, J=14.6, 10.8, 4.6 Hz, 2H).

Example S-4. Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(6-((4-((8R,9S)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-3-oxo-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-8-yl)phenyl)amino)-6-oxohexyl)benzamide(Compound ent-1.3)

Step-1: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-2-fluoro-N-(6-(4-(5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5yl)-3-oxo-3,7,8,9-tetrahydro-2H-pyrido[4,3,2-de]phthalazin-8-yl)phenylamino)-6-oxohexyl)benzamide

To a stirred solution of6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoicacid (0.210 g, 0.371 mmol) in DMF (8 mL) was added HATU (0.282 g, 0.743mmol, 2 eq) at 0° C. and the resulting mixture was stirred for 30 min.DIPEA (0.32 mL, 1.86 mmol, 5 eq) and8-(4-aminophenyl)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]phthalazin-3(7H)-one(PK-2) (0.17 g, 0.446 mmol, 1.2 eq) were then successively added to thereaction mixture and the resultant mixture was stirred at RT for 4 h.The reaction was monitored by TLC. Upon completion, the reaction mixturewas diluted with water (50 mL) and extracted using EtOAc (100 mL×2). Thecombined organic layer were washed with saturated NaHCO₃ solution (100mL), saturated NH₄Cl solution (100 mL), water (100 mL), brine (50 mL)dried over Na₂SO₄, filtered and concentrated under reduced pressure toafford a crude product which was purified by reverse phase HPLC toafford Compound ent-1.3. LC-MS 924 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD-d₄) δ8.15 (d, J=9.1 Hz, 2H), 7.98 (dd, J=8.1, 2.1 Hz, 1H), 7.90-7.79 (m, 2H),7.52 (d, J=8.9 Hz, 2H), 7.34 (d, J=8.4 Hz, 4H), 7.21 (dd, J=9.0, 2.5 Hz,1H), 6.90 (dd, J=10.8, 2.5 Hz, 1H), 3.59 (d, J=3.2 Hz, 3H), 3.46-3.36(m, 2H), 2.40 (t, J=7.4 Hz, 2H), 1.93 (s, 2H), 1.72 (dp, J=26.6, 7.3 Hz,4H), 1.60 (s, 6H), 1.48 (ddt, J=14.6, 10.8, 4.6 Hz, 2H).

Example S-5. Preparation of6-(((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)-N-(4-(5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-3-oxo-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-8-yl)phenyl)hexanamide(Compound 1.4)

Step-1: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(3 g, 10.3 mmol) in benzene (100 mL) was added PTSA (0.982 g, 5.1 mmol,0.5 eq) followed by addition of ethylene glycol (2.88 g, 51.61 mmol, 5eq) at RT. The resultant mixture was heated at 120° C. using Dean-Starkapparatus for 16 h. Reaction was monitored by TLC. Upon completion, thereaction mixture was diluted with water (200 mL) and extracted withEtOAc (350 mL). The organic layer was washed with saturated NaHCO₃solution (100 mL), water (200 mL), brine (100 mL) dried over Na₂SO₄,filtered and concentrated under reduced pressure to afford(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro spiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol. LC-MS 335 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 4.40 (d, J=5.26 Hz, 1H), 3.81 (s, 3H), 3.37-3.45 (m,2H), 1.82 (br. s., 2H), 1.70 (d, J=12.72 Hz, 2H), 1.59 (d, J=13.15 Hz,2H), 1.52 (d, J=10.52 Hz, 3H), 1.35 (br. s., 1H), 1.23-1.33 (m, 3H),1.03-1.21 (m, 5H), 0.87 (dt, J=7.89, 19.95 Hz, 4H), 0.76 (s, 3H), 0.61(s, 3H).

Step-2a: Preparation of 2-(6-chlorohexyloxy)tetrahydro-2H-pyran

To a stirred solution of 6-chlorohexan-1-ol (10 g, 73.158 mmol) indiethyl ether (100 mL) was added pTSA (0.05 g, 0.365 mmol, 0.005 eq)followed by addition of 3,4-dihydro-2H-pyran (8.4 g, 102.48 mmol, 1.4eq). The resulting reaction mixture was stirred at RT for 16 h. Thereaction was monitored by TLC. Upon completion, the reaction mixture wasdiluted with diethyl ether (350 mL). The organic layer was washed with20% KOH solution (100 mL), water (300 mL), brine (200 mL) dried overNa₂SO₄, filtered and concentrated under reduced pressure to afford2-(6-chlorohexyloxy)tetrahydro-2H-pyran. ¹H NMR (400 MHz, DMSO-d₆) δ4.50 (br. s., 1H), 4.25 (t, J=6.80 Hz, 2H), 4.20 (s, 1H), 4.00-4.08 (m,2H), 3.66-3.74 (m, 2H), 3.59 (dd, J=6.36, 16.01 Hz, 2H), 2.66 (s, 2H),1.81-1.90 (m, 2H), 1.67 (br. s., 1H), 1.38-1.45 (m, 2H), 1.20-1.27 (m,2H), 1.17 (t, J=7.02 Hz, 2H).

Step-2: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(6-(tetrahydro-2H-pyran-2-yloxy)hexyloxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane]

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol (2 g, 5.97 mmol) in xylene (50mL) was added NaNH₂ (50% suspension in toluene, 1.4 mL) and the mixturewas heated at 150° C. for 1 h. The reaction mixture was gradually cooledto RT, 2-(6-chlorohexyloxy) tetrahydro-2H-pyran (2.8 mL) was added to itand the resultant mixture was again heated to 150° C. for 16 h. Thereaction was monitored by TLC. Upon completion, the mixture was cooledto RT, quenched slowly with ice cold water (250 mL) and extracted withEtOAc (300 mL). The organic layer was washed with water (100 mL×2),brine (100 mL) dried over Na₂SO₄, filtered and concentrated underreduced pressure to obtain a crude product which was purified byCombiFlash chromatography (Teledyne Isco Rf+); compound eluting 30%EtOAc/hexane to afford(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(6-(tetrahydro-2H-pyran-2-yloxy)hexyloxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane].¹H NMR (400 MHz, CDCl3) δ 4.57 (d, J=3.95 Hz, 1H), 3.91-3.95 (m, 4H),3.82-3.91 (m, 2H), 3.73 (td, J=6.80, 9.65 Hz, 2H), 3.42-3.53 (m, 3H),3.33-3.42 (m, 3H), 3.26 (t, J=8.55 Hz, 2H), 1.91-2.01 (m, 2H), 1.75-1.90(m, 3H), 1.46-1.74 (m, 13H), 1.31-1.42 (m, 6H), 1.07-1.28 (m, 4H),0.83-0.97 (m, 4H), 0.81 (s, 4H), 0.70-0.76 (m, 4H), 0.07 (s, 3H), 0.00(s, 3H).

Step-3: Preparation of(5S,8R,9S,10S,13S,14S,17S)-17-(6-hydroxyhexyloxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(6-(tetrahydro-2H-pyran-2-yloxy)hexyloxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane](1 g, 1.92 mmol) in THE (25 mL) water (5 mL) was added 6N HCl (15 mL) atRT and the resultant reaction mixture was stirred at RT for 16 h. Thereaction was monitored by TLC. Upon completion, the reaction mixture wasdiluted with water (150 mL) and basified using with saturated NaHCO₃solution (pH ˜8). The aqueous layer was then extracted with EtOAc (200mL×3). The organic layer was washed with saturated NaHCO₃ solution (100mL), water (100 mL), brine (100 mL) dried over Na₂SO₄, filtered andconcentrated under reduced pressure to afford(5S,8R,9S,10S,13S,14S,17S)-17-(6-hydroxyhexyloxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-onewhich was used in the next step without further purification. ¹H NMR(400 MHz, CDCl3) δ 4.90 (br. s., 1H), 4.12 (d, J=7.45 Hz, 1H), 4.00 (br.s., 1H), 3.64 (t, J=6.58 Hz, 3H), 3.49-3.58 (m, 1H), 3.37-3.47 (m, 3H),3.27 (t, J=8.55 Hz, 1H), 2.22-2.43 (m, 3H), 2.10 (br. s., 1H), 1.93-2.07(m, 2H), 1.90 (d, J=11.84 Hz, 2H), 1.82 (d, J=7.89 Hz, 2H), 1.70 (d,J=10.09 Hz, 2H), 1.48 (d, J=4.38 Hz, 2H), 1.30-1.42 (m, 8H), 1.22-1.29(m, 2H), 1.01 (s, 3H), 0.77 (s, 3H).

Step-4: Preparation of6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoic acid

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-(6-hydroxyhexyloxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(0.1 g, 0.256 mmol) in acetone (5 mL) was added Jones reagent (0.5 mL)at 0° C. dropwise over a period of 20 min. The resultant mixture wasstirred at 0° C. for 5 min. The reaction was monitored by TLC. Aftercompletion, water (10 mL) was added and the resulting precipitate wasfiltered over Buchner funnel. The product obtained was washed with water(5 mL×2) and n-pentane (5 mL×2), dried under vacuum to afford6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid which was taken to next step without further purification. ¹H NMR(400 MHz, DMSO-d₆) δ 3.19-3.28 (m, 3H), 2.24-2.35 (m, 2H), 2.18 (t,J=7.24 Hz, 2H), 2.08 (d, J=13.59 Hz, 2H), 1.84-1.95 (m, 3H), 1.79 (d,J=10.52 Hz, 2H), 1.61 (d, J=13.59 Hz, 2H), 1.40-1.54 (m, 8H), 1.22-1.38(m, 8H), 1.19 (br. s., 1H), 1.09-1.18 (m, 2H), 0.97 (s, 3H), 0.69 (s,3H).

Step-5: Preparation of6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)-N-(4-(5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5yl)-3-oxo-3,7,8,9-tetrahydro-2H pyrido[4,3,2-de]phthalazin-8yl)phenyl)hexanamide

To a stirred solution of6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid (0.080 g, 0.197 mmol) in THE (5 mL) was added T₃P solution (50% inEtOAc, 0.25 mL, 0.435 mmol, 2.2 eq) at 0° C. and the resulting mixturewas stirred for 30 min. DIPEA (0.24 mL, 1.38 mmol, 7 eq)8-(4-aminophenyl)-5-fluoro-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]phthalazin-3(7H)-one(PK-1) (0.074 g, 0.197 mmol, 1 eq) were then successively added to thereaction mixture and the resultant mixture was stirred at RT for 16 h.The reaction was monitored by TLC. Upon completion, the reaction mixturewas diluted with water (50 mL) and extracted using EtOAc (100 mL×3). Thecombined organic layer were washed with saturated NaHCO₃ solution (80mL), saturated NH₄Cl solution (60 mL), water (80 mL), brine (60 mL)dried over Na₂SO₄, filtered and concentrated under reduced pressure toafford a crude product which was purified by reverse phase HPLC toafford Compound 1.4. LC-MS 764 [M+H]⁺. ¹H NMR (400 MHz, MeOD-d₄) δ 7.88(s, 1H), 7.56-7.49 (m, 2H), 7.35 (d, J=8.5 Hz, 2H), 7.21 (dd, J=9.0, 2.4Hz, 1H), 6.92 (dd, J=2.41, 10.74 Hz, 1H), 3.60 (s, 3H), 3.42-3.50 (m,3H), 2.31-2.40 (m, 2H), 1.91-2.12 (m, 5H), 1.88 (br. s., 2H), 1.64-1.78(m, 4H), 1.57 (dd, J=6.36, 13.81 Hz, 4H), 1.46 (dd, J=7.45, 15.79 Hz,4H), 1.13-1.40 (m, 10H), 0.82-0.97 (m, 4H), 0.71-0.82 (m, 3H).

Example S-6. Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(6-((4-(8-fluoro-1-oxo-2,3,4,6-tetrahydro-1H-azepino[5,4,3-cd]indol-5-yl)benzyl)amino)hexyl)benzamide(Compound 1.5)

Step-1: Preparation of4-(8-fluoro-6-oxo-3,4,5,6-tetrahydro-III-azepino[5,4,3-cd]indol-2-yl)benzaldehyde

To a stirred solution of2-bromo-8-fluoro-4,5-dihydro-1H-azepino[5,4,3-cd]indol-6(3H)-one (2 g,7.06 mmol, 1 eq) in DMA (37.5 mL) were added the solution of Na₂CO₃(1.48 g, 14.1 mmol, 2 eq) in H₂O (18.5 mL) and 4-formylphenylboronicacid (1.27 g, 8.4 mmol, 1.2 eq) at RT. The resulting mixture wasdegassed under nitrogen for 30 min at RT, followed by addition ofPd(dppf)Cl₂.DCM (0.142 g, 0.18 mmol, 0.025 eq). The mixture was furtherdegassed using nitrogen for 15 min. The reaction mixture was then heatedat 9.5° C. for 4 h. The reaction was monitored by TLC. Upon completion,the mixture was cooled to RT, water (30 mL) was added and the resultingprecipitate was filtered over Buchner funnel. The crude obtained waswashed with water (20 mL×2) and n-pentane (10 mL×2), dried under reducedpressure to afford4-(8-fluoro-6-oxo-3,4,5,6-tetrahydro-1H-azepino[5,4,3-cd]indol-2-yl)benzaldehyde.LC-MS 309 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.90 (s, 1H), 10.06 (s,1H), 8.31 (br. s., 1H), 8.02-8.08 (m, J=7.89 Hz, 2H), 7.81-7.89 (m,J=7.89 Hz, 2H), 7.46 (d, J=10.96 Hz, 1H), 7.37 (d, J=9.21 Hz, 1H), 3.41(br. s., 2H), 3.11 (br. s., 2H).

Step-2a: Preparation of tert-butyl 6-aminohexylcarbamate

To a stirred solution of hexane-1,6-diamine (10 g, 86.05 mmol) in DCM(400 mL) was added di-tert-butyl dicarbonate (4.69 g, 21.5 mmol, 0.25eq) dissolved in DCM (100 mL) dropwise over a period of 1 h and theresultant mixture was stirred at RT for 16 h. The reaction was thenfiltered over Buchner funnel to remove the unreacted hexane-1,6-diamine.The crude residue obtained was dissolved in water (200 mL) and thenextracted with DCM (200 mL×3). The combined organic layers were washedwith saturated NaHCO₃ solution (300 mL), water (200 mL), brine (150 mL)dried over Na₂SO₄, filtered and concentrated under reduced pressure toafford tert-butyl 6-aminohexylcarbamate which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ 4.70 (br, 1H),3.02 (m, 2H), 2.62 (m, 2H), 1.37 (s, 15H), 1.26 (m, 4H).

Step-2: Preparation of tert-butyl6-(4-(8-fluoro-6-oxo-3,4,5,6-tetrahydro-1H-azepino[5,4,3-cd]indol-2-yl)benzyl-amino)hexylcarbamate

To a stirred solution of4-(8-fluoro-6-oxo-3,4,_5,6-tetrahydro-1H-azepino[_5,4,3-cd]indol-2-yl)benzaldehyde(0.6 g, 1.94 mmol) in MeOH (36 mL) and THE (18 mL) was added tert-butyl6-aminohexylcarbamate (1.26 g, 5.8 mmol, 3 eq) and the reaction mixturewas stirred at RT for 2 h. NaBH₄ (0.22 g, 5.8 mmol, 3 eq) was thenslowly added to the mixture at 0° C. and the resulting mixture wasstirred at RT for 2 h. Reaction was monitored by TLC. Upon completion,the reaction mixture was diluted with water (100 mL) and extracted withEtOAc (150 mL×2). The combined organic layers were washed with water (75mL) and brine (75 mL) dried over Na₂SO₄, filtered and concentrated underreduced pressure to afford crude which was triturated using DCM/hexane(1:4) to afford tert-butyl6-(4-(8-fluoro-6-oxo-3,4,5,6-tetrahydro-1H-azepino[5,4,3-cd]indol-2-yl)benzyl-amino)hexylcarbamate.LC-MS 509 [M+H]⁺. ¹H NMR (400 MHz, MeOD-d₄) 7.60 (d, J=7.89 Hz, 2H),7.46-7.56 (m, 3H), 7.30 (br. s., 1H), 3.83 (s, 2H), 3.54 (br. s., 2H),3.14 (m, 3H), 3.02 (s, 1H), 2.62 (t, J=7.24 Hz, 2H), 1.57 (br. s., 4H),1.46 (m, 4H), 1.42 (s, 5H), 1.36 (br. s., 4H).

Step-3: Preparation of2-(4-((6-aminohexylamino)methyl)phenyl)-8-fluoro-4,5-dihydro-1H-azepino[5,4,3-cd]indol-6(3H)-one

To a stirred solution of tert-butyl6-(4-(8-fluoro-6-oxo-3,4,5,6-tetrahydro-1H-azepino[5,4,3-cd]indol-2-yl)benzyl-amino)hexylcarbamate(0.3 g, 0.58 mmol, 1 eq) in MeOH (1 mL) was added 4N-HCl in 1,4 dioxane(10 mL) at 0° C. The mixture was stirred at RT for 1 h. Reaction wasmonitored by TLC. Upon completion, the solvent is removed under reducedpressure to afford residue which was triturated using diethyl ether toafford2-(4-((6-aminohexylamino)methyl)phenyl)-8-fluoro-4,5-dihydro-1H-azepino[5,4,3-cd]indol-6(3H)-oneas a hydrochloride salt. LC-MS 445 [M+H]⁺.

Step-4: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-2-fluoro-N-(6-(4-(8-fluoro-6-oxo-3,4,5,6-tetrahydro-1H-azepino[5,4,3-cd]indol-2-yl)benzylamino)hexyl)benzamide

To a stirred solution of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoicacid (0.15 g, 0.32 mmol) in DMA (6 mL) was added HBTU (0.15 g, 0.39mmol, 1.2 eq) at 0° C. and the resulting mixture was stirred at sametemperature for 30 min. DIPEA (0.128 g, 0.99 mmol, 3 eq) and2-(4-((6-aminohexylamino)methyl)phenyl)-8-fluoro-4,5-dihydro-1H-azepino[5,4,3-cd]indol-6(3H)-one(0.221 g, 0.43 mmol, 1.5 eq) were then successively added to thereaction mixture and the resulting mixture was stirred at RT for 2 h.The reaction was monitored by TLC. Upon completion, the mixture wascooled to RT, water (30 mL) was added and the resulting precipitate wasfiltered over Büchner funnel. The crude obtained was purified by reversephase HPLC to afford Compound 1.5. LC-MS 842 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 11.67 (s, 1H), 8.50 (br. s., 1H), 8.40 (d, J=8.33 Hz, 1H),8.29 (s, 1H), 8.24 (br. s., 1H), 8.09 (s, 1H), 7.74 (s, 1H), 7.54-7.61(m, J=7.89 Hz, 2H), 7.44-7.51 (m, J=7.89 Hz, 2H), 7.40 (s, 1H), 7.43 (s,1H), 7.27-7.36 (m, 2H), 3.5-3.3 (m, 4H), 3.26 (d, J=5.70 Hz, 2H), 3.04(br. s., 2H), 1.89 (s, 2H), 1.48-1.58 (m, 6H), 1.47 (br. s., 2H), 1.33(br. s., 4H).

Example S-7. Preparation of4-[[4-fluoro-3-(piperazine-1-carbonyl)phenyl]methyl]-2H-phthalazin-1-one

Step-1: Preparation of (Z)-2-fluoro-5-((3-oxoisobenzofuran-1(3H)ylidene)methyl)benzonitrile

To a stirred solution of dimethyl 3-oxo-1,3-dihydroisobenzofuran-1-ylphosphonate (10 g, 41.28 mmol) and 2-fluoro-5-formylbenzonitrile (6.15g, 41.28 mmol, 1 eq) in THF (50 mL) was added triethylamine (5.76 mL,41.28 mmol, 1 eq) at 0° C. slowly. The resultant mixture was stirred atRT for 16 h. Reaction was monitored by TLC. Upon completion, water (200mL) was added and the resulting precipitate was filtered over Buchnerfunnel. The product obtained was washed with water (50 mL), hexanes (50mL), diethyl ether (30 mL), dried under vacuum afford to afford thetitle compound, as a mixture of E- and Z-isomers, which was used in nextstep without further purification. LC-MS 266 [M+H]+¹H NMR (400 MHz,CDCl₃) δ 8.13 (1H, m), 8.05 (1H, m), 7.98 (1H, m), 7.79 (2H, m), 7.61(1H, m), 7.30 (1H, m), 6.35 (1H, s).

Step-2: Preparation of(Z)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoic acid

To a stirred suspension of(Z)-2-fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)benzonitrile(3.7 g, 13.94 mmol) in water (20 mL) was added 13N NaOH solution (5 mL)and the mixture was heated under nitrogen at 90° C. for 16 h. Thereaction mixture was cooled to 70° C. and hydrazine hydrate (10 mL) wasadded and stirred for 16 hours at 70° C. Reaction was monitored by TLC.Upon completion, the reaction was cooled to RT and acidified using 2NHCl (pH 1-2) at 0-5° C. to obtain a precipitate. The precipitated solidwas filtered over Buchner funnel, washed with water (50 mL), n-hexanes(50 mL), diethyl ether (30 mL), dried under vacuum afford to afford thetitle compound. LC-MS 299 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 13.22 (1H,br s), 12.61 (1H, s), 8.27 (1H, m), 7.99-7.81 (4H, m), 7.59 (1H, m),7.25 (1H, m), 4.36 (2H, s).

Step-3: Preparation of tert-butyl4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1 yl)methyl)benzoyl)piperazine-1-carboxylate

To a stirred suspension of2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoic acid (0.100g, 0.335 mmol) in DMA (4 mL) was added HBTU (0.15 g, 0.402 mmol, 1.2 eq)at 0° C. and the mixture was stirred for 10 min. DIPEA (0.17 mL, 1.00mmol, 3 eq) and tert-butyl piperazine-1-carboxylate (0.075 g, 0.402mmol, 1.2 eq) were then successively added to the reaction mixture at 0°C. and the resultant reaction mixture was stirred at RT for 75 min.Reaction was monitored by TLC. After completion, water (10 mL) was addedand the resulting precipitate was filtered over Buchner funnel. Theproduct obtained was washed with water (10 mL×2) and n-pentane (10mL×2), dried under reduced pressure to afford the title compound whichwas taken to next step without further purification. LC-MS 467 [M+H]⁺.

Step-4: Preparation of4-[[4-fluoro-3-(piperazine-1-carbonyl)phenyl]methyl]-2H phthalazin-1-one

To tert-butyl4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazine-1-carboxylate(10.0 g, 76.23 mmol) was added 2N HCl in MeOH (30 mL) at RT and themixture was stirred at RT for 16 h. The reaction was monitored by ¹HNMR. Upon completion, the reaction mixture was concentrated underreduced pressure to afford compound, the compound was triturated withdiethylether and pentane afford the title compound as an hydrochloridesalt. LC-MS 403 [M+H]⁺. ¹H NMR (400 MHz, MeOD-d₄) δ 3.66 (s, 3H), 2.92(t, J=7.67 Hz, 2H), 2.37 (t, J=7.45 Hz, 2H), 1.62-1.71 (m, 6H),1.37-1.45 (m, 2H).

Example S-8. Preparation of (R)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropylsulfonyl)phenylamino)hexanoate

Step 1: Preparation of(R)-3-bromo-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

To a solution of (R)-3-bromo-2-hydroxy-2-methylpropanoic acid (13 g,71.04 mmol) in THF (250 mL) cooled to 0° C. was added SOCl₂ (14.1 g,118.53 mmol) dropwise and catalytic amount of DMF. The solution wasstirred at this temperature for 1.5 h. Then a solution of4-amino-2-(trifluoromethyl)-benzonitrile (7.4 g, 39.78 mmol) and TEA(15.8 g, 156.29 mmol) in THF (50 mL) was added dropwise at thistemperature. The mixture was allowed to warm to room temperatureovernight. TLC showed the reaction was completed. The mixture wasquenched with NaHCO₃ and extracted with EtOAc. The organic layer wasdried with Na₂SO₄ and concentrated under reduced pressure. The crudeproduct was purified by silica gel column to afford the title compound.

Step 2: Preparation of(R)-3-(4-aminophenylthio)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

To a solution of NaH (251 mg, 6.26 mmol) in THF (30 ml) was added4-aminobenzenethiol (712 mg, 5.68 mmol) in THF (20 mL) dropwise at 0° C.under N₂. The mixture was allowed to warm to room temperature andstirred for 1 h. Then a solution of(R)-3-bromo-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(2 g, 5.68 mmol) in THF (20 mL) was added dropwise to the mixture at 0°C. The mixture was stirred at room temperature overnight. TLC showed thereaction was completed. Water was added carefully and the mixture wasextracted with EtOAc, dried with Na₂SO₄ and concentrated under reducepressure. The compound was purified by silica gel column to afford thetitle compound.

Step 3: Preparation of (R)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropylthio)phenylamino)hexanoate

To a solution of(R)-3-(4-aminophenylthio)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(2 g, 5 mmol) in DMF (100 mL) was added K2CO₃ (2.1 g, 15 mmol), KI (100mg, 0.5 mmol), and methyl 6-bromohexanoate (10 g, 50 mmol). The mixturewas stirred at 60° C. overnight. TLC showed the reaction was completed.The mixture was washed with water, and extracted with EtOAc, dried overNa₂SO₄. The organic layer was concentrated under reduce pressure andpurified by flash chromatography to afford the title compound.

Step 4: Preparation of (R)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropylsulfonyl)phenylamino)hexanoate

To a solution of (R)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropylthio)phenylamino)hexanoate(2 g, 2.8 mmol) in DCM (60 mL) was added m-CPBA (1.83 g, 8.4 mmol), themixture was stirred at room temperature for 2 h. The mixture wasquenched with aq. NaHCO₃ and washed with water and extracted with DCM.The organic layer was dried over Na₂SO₄ and purified by flashchromatography to afford the title compound. ¹H NMR (400 MHz, DMSO-d₆) δ10.27 (s, 1H), 8.41 (s, 1H), 8.16 (d, J=8.6 Hz, 1H), 8.04 (d, J=8.6 Hz,1H), 7.45 (d, J=8.9 Hz, 2H), 6.63-6.37 (m, 3H), 6.30 (s, 1H), 3.79 (d,J=14.5 Hz, 1H), 3.58 (s, 3H), 3.47 (d, J=14.5 Hz, 1H), 2.89 (dq, J=11.8,7.0, 6.3 Hz, 2H), 2.31 (t, J=7.4 Hz, 2H), 1.55 (p, J=7.5 Hz, 2H), 1.47(q, J=7.2 Hz, 2H), 1.38 (s, 3H), 1.36-1.26 (m, 2H).

Example S-9. Preparation of (S)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropoxy)phenylamino)hexanoate

Step 1: Preparation of(S)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methyl-3-(4-nitrophenoxy)propanamide

To a stirring suspension of 4-nitrophenol (1.2 g, 8.55 mmol) and K2CO₃(3.9 g, 28.50 mmol) in isopropyl alcohol (40 mL) was added(R)-3-bromo-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(2 g, 5.70 mol). The reaction was refluxed for 2 h. TLC indicated thereaction was complete. The reaction mixture was concentrated in vacuo,diluted with water and extracted with EtOAc. The combined organic phaseswere washed with sat. NaHCO₃ and brine, dried over anhydrous Na₂SO₄ andconcentrated in vacuo to give the residue as an oil. The residue werepurified by column chromatography on silica gel to afford the titlecompound.

Step 2: Preparation of(S)-3-(4-aminophenoxy)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

To a stirring suspension of(S)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methyl-3-(4-nitrophenoxy)propanamide 3 (2.0 g, 4.64 mmol) in EtOH (20 mL) and H₂O (20 mL) wasadded NH₄Cl (2.5 g, 46.42 mmol) and iron powder (2.1 g, 3.71 mmol). Thereaction was degassed with N₂ and stirred at 90° C. for 1 h. TLCindicated the reaction was complete. The mixture was diluted with EtOAc,filtered off with a pad of Celite, washed with brine, dried overanhydrous Na₂SO₄ and concentrated in vacuo to afford the title compound.

Step 3: Preparation of (S)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropoxy)phenylamino)hexanoate

To a solution of(S)-3-(4-aminophenoxy)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(1.7 g, 4.48 mmol) and methyl 6-bromohexanoate (0.7 g, 3.45 mmol) in DMF(30 mL) was added K2CO₃ (1.0 g, 7.56 mmol) and KI (0.6 g, 3.80 mmol).The reaction was stirred at 100° C. for 4 h. TLC indicated ˜30% of(S)-3-(4-aminophenoxy)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamidewas still remained. The mixture was diluted with water, extracted withEtOAc. The combined organic phases were washed with brine, dried overanhydrous Na₂SO₄ and concentrated in vacuo to give the residue. Theresidue was purified by column chromatography on silica gel to affordthe title compound. LCMS 508.6 [M+1]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 10.54(s, 1H), 8.57 (d, J=2 Hz, 1H), 8.32 (dd, J=2 Hz, 8.4 Hz 1H), 8.11 (d,J=8.8 Hz, 1H), 6.70-6.68 (m, 1H), 6.48-6.46 (m, 1H), 6.17 (s, 1H), 5.18(s, 1H), 4.10 (d, J=9.6 Hz, 1H), 3.87 (d, J=9.6 Hz, 1H), 3.58 (s, 1H),2.32 (t, J=7.2 Hz), 1.57-1.49 (m, 4H), 1.41 (s, 3H), 1.36-1.33 (m, 2H).

Example S-10. Preparation of(R)-3-(4-(6-bromohexyloxy)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

Step 1: Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-3-(4-hydroxyphenylthio)-2-methylpropanamide

A solution of(R)-3-bromo-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(1.5 g, 4.27 mmol), 4-mercaptophenol (0.539 g, 4.27 mmol) and K2CO₃(0.649 g, 4.69 mmol) in acetone (15 mL). The reaction mixture wasstirred at rt for 3 h. TLC showed the reaction mixture was complete. Thesolution was concentrated in vacuo. The crude product was purified bycolumn chromatography to afford the title compound.

Step 2: Preparation of(R)-3-(4-(6-bromohexyloxy)phenylthio)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

A mixture of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-3-(4-hydroxyphenylthio)-2-methylpropanamide(1.34 g, 3.38 mmol), 1,6-dibromohexane (8.25 g, 33.81 mmol) and K2CO₃(2.34 g, 16.9 mmol) in ACN (20 mL). The reaction mixture was stirred atrt for 3 h. TLC showed the mixture was complete. The solution wasconcentrated in vacuo, and purified by column chromatography to affordthe title compound.

Step 3: Preparation of(R)-3-(4-(6-bromohexyloxy)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

A mixture of(R)-3-(4-(6-bromohexyloxy)phenylthio)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(1.5 g, 2.68 mmol) in DCM (15 mL) was cooled to 0° C., and m-CPBA (1.39g, 8.04 mmol) was added. The reaction mixture was stirred at rt for 2 h.TLC showed the mixture was complete. The solution was poured into 1NNaOH and extracted with DCM (50 mL×3). The organic layer was washed withbrine, dried over Na₂SO₄, concentrated and purified by columnchromatography to afford the title compound. ¹H NMR (400 MHz, CDCl₃) δ8.94 (s, 1H), 7.88 (d, J=2.8 Hz 1H), 7.74-7.68 (m, 2H), 7.34-7.30 (m,2H), 6.66-6.62 (m, 2H), 3.81-3.71 (m, 4H), 3.43 (t, J=6.8 Hz 2H), 2.99(d, J=14.4 Hz 1H), 1.93-1.86 (m, 2H), 1.78-1.71 (m, 2H), 1.57-1.43 (m,7H).

Example S-11. Preparation of4-(1-(4-(2-bromoethoxy)phenyl)-2-phenylbut-1-enyl)phenyl pivalate

Step 1: Preparation of 4-(4-hydroxybenzoyl)phenyl

Part 1: To a solution of bis(4-hydroxyphenyl)methanone 1 (2 g, 9.34mmol) in 100 mL of THF was added Et₃N (3.8 g, 37.36 mmol). Then PivCl(2.82 g, 23.35 mmol) was added dropwise to the mixture at 0° C. Themixture was allowed to wane to rt and stirred overnight. The reactionmixture was quenched with water and extracted with EtOAc. The organiclayers were washed with 1N HCl and brine, dried over Na₂SO₄,concentrated under reduced pressure to get a crude product.

Part 2: To a solution of the crude product (3.5 g) in THF (45 mL) andMeOH (3 mL) was added LiOH (290 mg, 12.14 mmol), the mixture was stirredat rt overnight. The reaction mixture was concentrated under reducedpressure and purified by silica gel chromatography to afford the titlecompound.

Step 2: Preparation of (E)-4-(1-(4-hydroxyphenyl)-2phenylbut-1-enyl)phenyl pivalate

To a suspension of Zn (1.7 g, 26.8 mmol) in anhydrous THF (150 mL) underN₂ was added TiCl₄ (2.5 g, 13.4 mmol) at 0° C. The mixture was refluxedfor 2 h and cooled at 40° C. A mixture of 4-(4-hydroxybenzoyl)phenylpivalate (1 g, 3.35 mmol) and propiophenone (1.5 g, 10.72 mmol) inanhydrous THF (50 mL) was added at once and the mixture was refluxed for1 h. The reaction mixture was quenched with 10% K2CO₃, extracted withEtOAc, washed with brine, dried over Na₂SO₄, purified by silica gelchromatography to get a crude product, then added MeOH (10 mL) andstirred for 20 min to obtain the title compound.

Step 3: Preparation of4-(1-(4-(2-bromoethoxy)phenyl)-2-phenylbut-1-enyl)phenyl pivalate

To a solution of (E)-4-(1-(4-hydroxyphenyl)-2-phenylbut-1-enyl)phenylpivalate 4 (0.4 g, 1 mmol) in acetone (6 mL) was added K2CO₃ (0.56 g, 4mmol), 1,2-dibromoethane 5 (1.9 g, 10 mmol). The mixture was stirred at110° C. overnight. TLC showed the reaction was completed. The mixturewas washed with water, and extracted with EtOAc, dried over Na₂SO₄. Theorganic layer was concentrated under reduced pressure and purified bysilica gel column to afford the title compound. ¹H NMR (400 MHz, CDCl₃,E/Z=1:1) δ 7.23 (d, J=8.7 Hz, 2H), 7.20-7.14 (m, 6H), 7.14-7.07 (m, 6H),7.05 (d, J=8.6 Hz, 2H), 6.87 (dd, J=15.3, 8.8 Hz, 4H), 6.77 (d, J=8.9Hz, 2H), 6.70 (d, J=8.8 Hz, 2H), 6.55 (d, J=8.9 Hz, 2H), 4.31 (t, J=6.3Hz, 2H), 4.15 (t, J=6.3 Hz, 2H), 3.66 (t, J=6.3 Hz, 2H), 3.55 (t, J=6.3Hz, 2H), 2.57-2.39 (m, 4H), 1.37 (s, 9H), 1.28 (s, 9H), 0.97-0.89 (m,6H).

Example S-12. Preparation of(R)-3-(4-(2-bromoethylamino)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)-phenyl)-2-hydroxy-2-methylpropanamide

Step 1: Preparation of(R)-3-(4-aminophenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

To a solution of(R)-3-(4-aminophenylthio)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide1 (3.0 g, 7.59 mmol) in DCM (20 mL) remained at 0° C. was added 85%mCPBA (4.6 g, 22.76 mmol). The reaction was stirred at rt for 4 h. TLCindicated the reaction was complete. The mixture was washed with sat.NaHCO₃. The organic phase was washed with brine, dried over anhydrousNa₂SO₄, concentrated under reduced pressure and purified by columnchromatography to afford the title compound.

Step 2: Synthesis of 2-bromoacetaldehyde

A mixture of 2-bromo-1,1-diethoxyethane (20 g, 0.101 mol) in HBr (47%,32 mL) was stirred at 100° C. for 2 h. The mixture was extracted withEt₂O. The combined organic layers were washed with brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to give thetitle compound, which was used in the next step directly.

Step 3: Synthesis of(R)-3-(4-(2-bromoethylamino)phenylsulfonyl)-JV-(4-cyano-3-(trifluoromethyl)-phenyl)-2-hydroxy-2-methylpropanamide

To a solution of(R)-3-(4-aminophenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(2 g, 4.68 mmol) and 2-bromo-1,1-diethoxyethane (5.2 g, 42.11 mmol) inMeOH (40 mL) was added AcOH (0.18 g, 3.04 mmol) followed by BH₃NMe₃ (4.8g, 65.51 mmol). The reaction was monitored by TLC and additionalportions of 2-bromo-1,1-diethoxyethane was added until the conversionwas complete. The mixture was diluted with EtOAc, washed with sat.NaHCO₃ and brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure. The crude material was purified by columnchromatography to afford the title compound. LCMS 536.1 [M+1]⁺. ¹H NMR(600 MHz, DMSO-d₆) δ 10.33 (s, 1H), 8.44 (d, J=3 Hz, 1H), 8.20 (dd, J=3Hz, 12.6 Hz, 1H), 8.08 (d, J=12.6 Hz, 1H), 7.51 (d, J=13.8 Hz, 2H), 6.60(d, J=13.8 Hz, 2H), 6.33 (s, 1H), 2.98 (s, 3H), 3.82 (d, J=21.6 Hz, 1H),3.53 (m, 6H), 1.39 (s, 3H).

Example S-13. Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide(Compound 1.68)

Step-A: Preparation of(S)-6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropylsulfonyl)phenylamino)hexanoicacid

To (5)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropylsulfonyl)phenylamino)hexanoate (0.20 g, 0.360 mmol) in THF:MeOH:H₂O (6mL:3 mL:1 mL) was added lithium hydroxide monohydrate (0.151 mg, 3.6mmol, 10 eq) and the mixture was heated at 50° C. for 2 h. Aftercompletion, the mixture was concentrated under reduced pressure. Theresidue obtained was acidified with 1N HCl (pH ˜2) to obtain aprecipitate which was filtered over Buchner funnel to afford the titlecompound. LC-MS 542 [M+H]⁺.

Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1 yl)-6-oxohexylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide (Compound 1.68)

To a stirred solution of(5)-6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropylsulfonyl)phenylamino)hexanoicacid (0.20 g, 0.369 mmol) in DMF (5 mL) was added HATU (0.21 g, 0.554mmol, 1.5 eq) at 0° C. and the mixture was stirred at 0° C. for 30 min.DIPEA (0.34 mL, 1.84 mmol, 5 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride salt (0.297 g, 0.739 mmol, 2.0 eq) were then successivelyadded to the mixture and the resultant mixture was stirred at RT for 16h. The reaction was monitored by TLC and LC-MS. After completion, thereaction was diluted with EtOAc (250 mL). The organic layer was washedwith water (100 mL), brine (50 mL) dried over Na₂SO₄, filtered andconcentrated under reduced pressure to obtain a crude residue which waspurified by reversed phase HPLC to afford Compound 1.68. LC-MS 890[M+H]⁺. ¹H NMR (400 MHz, CD₃OD-d₄) δ 8.36 (d, J=7.8 Hz, 1H), 8.21 (s,1H), 7.98-7.77 (m, 5H), 7.56-7.43 (m, 3H), 7.38 (d, J=6.4 Hz, 1H), 7.15(t, J=9.0 Hz, 1H), 6.42 (dd, J=9.0, 5.2 Hz, 2H), 4.38 (s, 2H), 4.00 (d,J=14.5 Hz, 1H), 3.80-3.60 (m, 4H), 3.55-3.38 (m, 3H), 3.28 (s, 1H), 2.89(tt, J=12.3, 6.0 Hz, 2H), 2.43 (dt, J=28.6, 7.4 Hz, 2H), 1.60 (ddt,J=36.4, 15.2, 7.6 Hz, 4H), 1.44 (d, J=14.4 Hz, 5H).

Example S-14. Preparation of(S)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexylamino)phenoxy)-2-hydroxy-2-methylpropanamide(Compound 1.83)

Step-A: Preparation of(R)-6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropoxy)phenylamino)hexanoicacid

To (R)-methyl6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropoxy)phenylamino)hexanoate(0.20 g, 0.360 mmol) in THF:MeOH:H₂O (6 mL:3 mL:1 mL) was added lithiumhydroxide monohydrate (0.151 mg, 3.6 mmol, 10 eq)) and the mixture washeated at 50° C. for 2 h. After completion, the mixture was concentratedunder reduced pressure. The residue obtained was acidified with 1N-HCl(pH˜2) to obtain a precipitate which was filtered over Buchner funnel toafford the title compound. LC-MS 494 [M+H]⁺.

Preparation of(S)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexylamino)phenoxy)-2-hydroxy-2-methylpropanamide (Compound 1.83)

To a stirred solution of(R)-6-(4-(3-(4-cyano-3-(trifluoromethyl)phenylamino)-2-hydroxy-2-methyl-3-oxopropoxy)phenylamino)hexanoicacid (0.20 g, 0.405 mmol) in DMF (5 mL) was added HATU (0.23 g, 0.608mmol, 1.5 eq) at 0° C. and the mixture was stirred at 0° C. for 30 min.DIPEA (0.37 mL, 2.02 mmol, 5 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.24 g, 0.608 mmol, 1.5 eq) were then successively addedto the reaction mixture and the mixture was stirred at RT for 16 h. Thereaction was monitored by TLC and LC-MS. After completion, the mixturewas diluted with EtOAc (250 mL). The organic layer was washed with water(100 mL), brine (50 mL) dried over Na₂SO₄, filtered and concentratedunder reduced pressure to obtain a crude residue which was purified byreversed phase HPLC to afford Compound 1.83. LC-MS 842.8 [M+H]⁺. ¹H NMR(400 MHz, DMSO-d₆) δ 8.55 (d, J=2.9 Hz, 1H), 8.28 (d, J=10.5 Hz, 2H),8.08 (d, J=8.6 Hz, 1H), 7.96 (d, J=8.1 Hz, 1H), 7.89 (t, J=7.4 Hz, 1H),7.83 (t, J=7.5 Hz, 1H), 7.44 (t, J=6.6 Hz, 1H), 7.36 (s, 1H), 7.23 (t,J=9.0 Hz, 1H), 7.06 (d, J=8.5 Hz, 1H), 6.89 (s, 1H), 6.67 (d, J=7.8 Hz,2H), 6.45 (d, J=8.1 Hz, 2H), 5.06 (t, J=5.5 Hz, 1H), 4.33 (s, 2H), 4.06(d, J=9.5 Hz, 1H), 3.86 (t, J=9.9 Hz, 1H), 3.62 (s, 2H), 3.51 (t, J=4.9Hz, 2H), 3.15 (d, J=14.4 Hz, 2H), 2.91 (d, J=17.5 Hz, 2H), 2.28 (d,J=7.9 Hz, 1H), 1.69 (s, 2H), 1.51 (q, J=6.5, 5.7 Hz, 4H), 1.38 (d,J=12.8 Hz, 6H).

Example S-15. Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)hexyloxy)phenylsulfonyl)-2-hydroxy-2-methylpropanamide(Compound 1.96)

To a stirred solution of4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.15 g, 0.254 mmol) in EtOH (4 mL) was added DIPEA (0.234mL, 1.27 mmol, 5 eq) followed by the addition of(S)-3-(4-(6-bromohexyloxy)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(0.153 g, 0.381 mmol, 2.0 eq) and the mixture was stirred at 80° C. for16 h. The reaction was monitored by TLC and LC-MS. After completion, thereaction was concentrated under reduced pressure to obtain a cruderesidue which was purified by reversed phase HPLC to afford Compound1.96. LC-MS 877 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 12.53 (d, J=39.4 Hz,1H), 10.47 (s, 1H), 8.39 (d, J=2.1 Hz, 1H), 8.26 (d, J=7.8 Hz, 1H),8.18-8.10 (m, 1H), 8.05 (d, J=8.6 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.88(t, J=7.5 Hz, 1H), 7.82 (t, J=7.4 Hz, 1H), 7.73 (d, J=8.4 Hz, 2H), 7.42(ddd, J=8.2, 5.1, 2.4 Hz, 1H), 7.30 (dd, J=6.4, 2.4 Hz, 1H), 7.21 (t,J=9.0 Hz, 1H), 6.94 (d, J=8.5 Hz, 2H), 4.32 (s, 2H), 3.85 (dt, J=15.8,11.9 Hz, 3H), 3.65-3.56 (m, 3H), 3.13 (t, J=5.0 Hz, 2H), 2.37 (t, J=5.0Hz, 2H), 2.27 (t, J=7.3 Hz, 2H), 2.24-2.17 (m, 2H), 1.78 (s, 2H), 1.66(p, J=7.2 Hz, 2H), 1.50-1.27 (m, 7H).

Example S-16. Preparation of(S)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(2-(4-((4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)ethylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide(Compound 1.97)

Step-1: Preparation of(S)-3-(4-(2-azidoethylamino)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide

To a stirred solution of(S)-3-(4-(2-bromoethylamino)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide (0.30 g, 0.56 mmol) in DMF (5 mL)was added sodium azide (0.073, 0.112 mmol, 2 eq)) at RT and the mixturewas heated at 80° C. for 2 h. The reaction was monitored by TLC. Uponcompletion, the mixture was diluted with EtOAc (250 mL). The organiclayer was washed with water (100 mL×2), brine (100 mL), dried withanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toobtain a crude which was purified by CombiFlash chromatography to affordthe title compound. LC-MS 497 [M+H]⁺.

Step-2a: Preparation of 4-(4-fluoro-3-(4-(prop-2ynyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.40 g, 0.99 mmol) in EtOH (5 mL) was added DIPEA (0.91mL, 4.97 mmol, 5 eq) followed by the addition of bromoethyne (0.176 g,1.49 mmol, 2.0 eq) and the mixture was heated at 80° C. for 16 h. Thereaction was monitored by TLC and LC-MS. After completion, the mixturewas concentrated under reduced pressure to obtain a crude which waspurified by CombiFlash chromatography to afford the title compound.LC-MS 405 [M+H]⁺.

Step-2: Preparation of(S)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(2-(4-((4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1yl)methyl)-1H-1,2,3-triazol-1-yl)ethylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide(Compound 1.97)

To a stirred solution of(S)-3-(4-(2-azidoethylamino)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide (0.150 g, 0.301 mmol) and4-(4-fluoro-3-(4-(prop-2-ynyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one (0.182 g, 0.452 mmol,15 eq) in MeCN:H₂O (2:1) (6 mL) were successively added cupper suphatepentahydrate (0.024 g, 0.15 mmol, 0.5 eq) and hydrazine hydrate (0.011mg, 1.00 mmol, 1 eq) and the resultant mixture was heated at 60° C. for2 h. The reaction was monitored by TLC and LC-MS. After completion, themixture was diluted with water (50 mL) and extracted with ethylacetate(50 mL×2). The combined organic layers were washed with 10% NH₄OHsolution, dried with anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to obtain a crude which was purified by CombiFlashchromatography to afford Compound 1.97. LC-MS 901 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 12.59 (s, 1H), 10.35 (s, 1H), 8.43 (d, J=2.1 Hz, 1H),8.29-8.15 (m, 2H), 8.06 (d, J=8.6 Hz, 1H), 8.00 (s, 1H), 7.96 (d, J=8.0Hz, 1H), 7.88 (t, J=7.5 Hz, 1H), 7.82 (t, J=7.5 Hz, 1H), 7.48 (d, J=8.5Hz, 2H), 7.42 (s, 1H), 7.32 (d, J=6.1 Hz, 1H), 7.22 (t, J=8.9 Hz, 1H),6.72 (t, J=6.0 Hz, 1H), 6.56 (d, J=8.5 Hz, 2H), 6.32 (s, 1H), 4.47 (s,2H), 4.32 (s, 2H), 3.78 (d, J=14.5 Hz, 1H), 3.52 (dt, J=19.4, 11.7 Hz,6H), 3.14 (s, 2H), 2.40 (s, 2H), 2.28 (s, 1H), 1.38 (s, 3H), 1.23 (s,1H).

Example S-17. Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-2-oxoethylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide(Compound 1.98)

Step-1: Preparation of4-(3-(4-(2-bromoacetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To 4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (1.0 g, 2.48 mmol) in DCM (5 mL) was added Et₃N (3.58 mL,24.8 mmol) at RT and the mixture was slowly added to a previouslystirred solution of 2-bromoacetyl chloride (3.85 g, 24.8 mmol) in DCM(20 mL) at 0° C. The resultant mixture was stirred at RT for 10 min andmonitored by TLC and LC-MS. After completion, the reaction was quenchedwith saturated NaHCO₃ solution (30 mL) and extracted with DCM (200mL×2). The combined organic layers were washed with H₂O (100 mL), brine(50 mL), dried over anhydrous sodium sulphate and concentrated underreduced pressure to obtain a crude residue which was purified byCombiFlash chromatography to afford the title compound. LC-MS 487 [M+H]⁺

Step-2: Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1 yl)-2-oxoethylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide

To a stirred solution of(R)-3-(4-aminophenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(0.105 g, 0.246 mmol, 1 eq) in DMF (5 ml) was added4-(3-(4-(2-bromoacetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one(0.12 g, 0.246 mmol, 1 eq) and the reaction mixture was stirred at 80°C. for 16 h. The reaction was monitored by TLC and LC-MS. Uponcompletion, the mixture was diluted with EtOAc (100 mL). The organiclayer was washed with ice-cold H₂O (50 mL×2), brine (30 mL), dried withanhydrous sodium sulphate and concentrated under reduced pressure toobtain a crude residue which was purified by reversed phase HPLC toafford the title compound. LC-MS: 487 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ12.60 (s, 1H), 10.33 (s, 1H), 8.43 (d, J=2.1 Hz, 1H), 8.31-8.23 (m, 1H),8.21-8.15 (m, 1H), 8.06 (d, J=8.5 Hz, 1H), 7.97 (t, J=5.2 Hz, 1H), 7.90(d, J=6.6 Hz, 1H), 7.85 (d, J=7.9 Hz, 1H), 7.48 (t, J=9.7 Hz, 1H),7.42-7.33 (m, 3H), 7.25 (q, J=7.6 Hz, 2H), 6.64 (dd, J=11.7, 8.2 Hz,2H), 6.32 (s, 1H), 4.34 (s, 2H), 3.90 (d, J=19.0 Hz, 2H), 3.79 (d,J=14.5 Hz, 1H), 3.70 (s, 1H), 3.63 (s, 1H), 3.57 (s, 2H), 3.52 (s, 1H),3.42 (s, 2H), 3.26 (s, 2H), 3.20 (s, 1H), 1.39 (s, 3H).

Example S-18. Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)ethylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide (Compound 1.99)

Step-1: Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1 yl)ethylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide

To a stirred solution of4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.075 g, 0.187 mmol, 1.0 eq) in EtOH (10 mL) was addedDIPEA (0.201 mL, 1.12 mmol, 6 eq) followed by the addition of(R)-3-(4-(2-bromoethylamino)phenylsulfonyl)-N-(4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide(0.10 g, 0.187 mmol) and the mixture was stirred at 80° C. for 16 h. Thereaction was monitored by TLC and LC-MS. After completion, the reactionwas concentrated under reduced pressure to obtain a crude residue whichwas purified by reversed phase HPLC to afford the title compound.LC-MS=820 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 12.61 (br. s., 1H), 8.41(br. s., 1H), 8.26 (d, J=7.5 Hz, 1H), 8.14 (d, J=7.0 Hz, 1H), 8.03 (d,J=7.9 Hz, 1H), 7.96 (d, J=7.5 Hz, 1H), 7.91-7.80 (m, 2H), 7.50-7.38 (m,3H), 7.32 (d, J=4.4 Hz, 1H), 7.22 (t, J=9.0 Hz, 1H), 6.58-6.48 (m, 2H),6.40 (br. s., 1H), 3.76 (d, J=14.0 Hz, 1H), 3.62 (br. s., 2H), 3.50 (d,J=14.5 Hz, 1H), 3.16 (br. s., 2H), 3.07 (d, J=5.3 Hz, 2H), 2.49-2.4 (m,4H), 2.31 (br. s., 2H), 1.68 (s, 2H), 1.37 (s, 3H).

Example S-19. Preparation of4-(3-(4-(2-(2-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)ethoxy)acetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one(Compound 1.104)

Step-1: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(15 g, 51.65 mmol) in benzene (500 mL) was added PTSA (4.85 g, 25.8mmol, 0.5 eq) followed by addition of ethylene glycol (14.5 mL, 258mmol, 5 eq) at RT. The resultant mixture was refluxed using Dean-Starkapparatus for 16 h. The reaction was monitored by TLC. Upon completion,the reaction mixture was diluted with water (500 mL) and extracted withEtOAc (500 mL×2). The combined organic layers were washed with saturatedNaHCO₃ solution (250 mL), water (400 mL), brine (200 mL) dried overNa₂SO₄, filtered and concentrated under reduced pressure to afford thetitle compound. ¹HNMR (400 MHz, DMSO-d₆) δ 4.40 (d, J=5.26 Hz, 1H), 3.81(s, 3H), 3.45-3.37 (m, 2H), 1.82 (br. s., 2H), 1.70 (d, J=12.72 Hz, 2H),1.59 (d, J=13.15 Hz, 2H), 1.52 (d, J=10.52 Hz, 3H), 1.35 (br. s., 1H),1.33-1.23 (m, 3H), 1.21-1.03 (m, 5H), 0.87 (dt, J=7.89, 19.95 Hz, 4H),0.76 (s, 3H), 0.61 (s, 3H)

Step-2a: Preparation of 2-(2-(2-chloroethoxy)ethoxy)tetrahydro-2H-pyran

To a stirred solution of 2-(2-chloroethoxy)ethanol (10 g, 80.6 mmol) indiethyl ether (150 mL) was added PTSA (1.54 g, 0.806 mmol, 0.1 eq)followed by addition of 3,4-dihydro-2H-pyran (8.12 g, 96.7 mmol, 1.2eq). The resulting reaction mixture was stirred at RT for 16 h. Thereaction was monitored by TLC. Upon completion, the reaction mixture wasdiluted with diethyl ether (700 mL). The organic layer was washed with20% KOH solution (300 mL), water (600 mL), brine (300 mL) dried overNa₂SO₄, filtered and concentrated under reduced pressure to afford thetitle compound which was taken to next step without furtherpurification. ¹HNMR (400 MHz, DMSO-d₆) δ 4.62-4.56 (m, 1H), 3.75-3.66(m, 6H), 3.63-3.56 (m, 2H), 3.53-3.46 (m, 1H), 3.46-3.39 (m, 1H),1.75-1.67 (m, 1H), 1.65-1.57 (m, 1H), 1.52- 1.39 (m, 4H).

Step-2: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-(2-(tetrahydro-2Hpyran-2-yloxy)ethoxy)ethoxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane]

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol(1.2 g, 3.59 mmol) in xylene (12 mL) was added NaNH₂ (50% suspension intoluene, 3.5 mL) and the mixture was heated at 150° C. for 1 h. Thereaction mixture was gradually cooled to 25° C.,2-(6-chlorohexyloxy)tetrahydro-2H-pyran (7.47 g, 35.9 mmol) was added toit and the resultant mixture was further heated at 150° C. for 16 h. Thereaction was monitored by TLC. Upon completion, the mixture was cooledto RT, quenched slowly with ice cold water (500 mL) and extracted withEtOAc (300 mL×2). The combined organic layer were washed with water (250mL×2), brine (200 mL) dried over Na₂SO₄, filtered and concentrated underreduced pressure to obtain a crude product which was purified byCombiFlash chromatography to afford the title compound. ¹HNMR (400 MHz,CDCl3) δ 4.64 (t, J=3.5 Hz, 1H), 3.93 (s, 4H), 3.90-3.81 (m, 4H), 3.74(dd, J=5.5, 3.7 Hz, 1H), 3.70-3.64 (m, 4H), 3.64-3.54 (m, 4H), 3.54-3.44(m, 2H), 3.32 (t, J=8.3 Hz, 2H), 2.02-1.93 (m, 2H), 1.90-1.80 (m, 2H),1.66-1.60 (m, 6H), 1.44-1.33 (m, 4H), 1.28-1.16 (m, 6H), 0.98- 0.84 (m,4H), 0.84-0.77 (m, 2H), 0.76-0.69 (m, 2H).

Step-3: Preparation of(5S,8R,9S,10S,13S,14S,17S)-17-(2-(2-hydroxyethoxy)ethoxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-(2-(tetrahydro-2H-pyran-2-yloxy)ethoxy)ethoxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane](0.5 g, 0.988 mmol) in THF (30 mL)-H₂O (10 mL) was added 6N TLC (20 mL)at RT and the resultant reaction mixture was stirred at RT for 16 h. Thereaction was monitored by TLC. Upon completion, the reaction mixture wasdiluted water (30 mL) and basified using with saturated NaHCO₃ solution(pH ˜8). The aqueous layer was then extracted with EtOAc (40 mL×3). Theorganic layer was washed with saturated NaHCO₃ solution (30 mL), water(30 mL), brine (20 mL) dried over Na₂SO₄, filtered and concentratedunder reduced pressure to afford the title compound which was used inthe next step without further purification. ¹HNMR (400 MHz, CDCl3) δ3.75-3.69 (m, 2H), 3.69-3.60 (m, 4H), 3.36 (t, 0.1=8.3 Hz, 1H), 2.66 (t,J=6.4 Hz, 1H), 2.37 (dd, J=13.8, 6.4 Hz, 1H), 2.33-2.25 (m, 2H),2.12-2.08 (m, 1H), 2.07-1.96 (m, 2H), 1.96-1.87 (m, 2H), 1.70 (dd,J=13.4, 3.3 Hz, 2H), 1.47-1.40 (m, 2H), 1.27-1.28 (m, 4H), 1.27-1.15 (m,4H), 1.04-0.99 (m, 4H), 0.97 (d, J=7.9 Hz, 2H), 0.88-0.81 (m, 2H), 0.79(s, 2H).

Step-4: Preparation of2-(2-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)ethoxy) acetic acid

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-(2-(2-hydroxyethoxy)ethoxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(0.40 g, 0.947 mmol) in acetone (2.5 mL) was added Jones reagent (2.4mL) at 0° C. dropwise over a period of 30 min. The resultant mixture wasstirred at 0° C. for 10 min. The reaction was monitored by TLC. Aftercompletion, water (100 mL) was added and the resulting precipitate wasfiltered over Buchner funnel. The product obtained was washed with water(50 mL×2) and n-pentane (50 mL×2), dried under vacuum to afford thetitle compound which was taken to next step without furtherpurification. ¹HNMR (400 MHz, DMSO-d₆) δ 12.51 (br. s., 1H), 4.02 (s,2H), 3.58-3.46 (m, 3H), 2.39 (dd, J=6.8, 14.7 Hz, 2H), 2.34-2.23 (m,2H), 2.08 (d, J=13.6 Hz, 2H), 1.97-1.85 (m, 3H), 1.80 (d, J=12.7 Hz,1H), 1.61 (d, J=13.2 Hz, 1H), 1.51 (br. s., 3H), 1.42-1.33 (m, 2H),1.32-1.06 (m, 8H), 1.00-0.92 (m, 2H), 0.70 (s, 2H)

Step-5: Preparation of 9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)ethoxy)acetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To a stirred solution of2-(2-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)ethoxy)aceticacid (0.05 g, 0.127 mmol) in DMF (5 mL) was added HATU (0.058 g, 0.153mmol, 1.2 eq) at 0° C. and the resulting reaction mixture was stirredfor 10 min. DIPEA (0.114 mL, 0.637 mmol, 5 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.035 g, 0.089 mmol, 0.7 eq) were then successively addedto the mixture and the mixture was stirred at RT for 2 h. The reactionwas monitored by TLC & LC-MS. After completion, water (10 mL) was addedand the resulting precipitate was filtered over Buchner funnel. Theproduct obtained was washed with water (25 mL×2) and n-pentane (25mL×2), dried under vacuum to obtain a crude residue which was purifiedby reversed phase HPLC to afford the title compound. LC-MS 741 [M+H]⁺.¹HNMR (400 MHz, MeOD-d₄) δ 8.37 (d, J=7.8 Hz, 1H), 7.96 (t, J=7.5 Hz,1H), 7.86 (dt, J=18.5, 7.4 Hz, 2H), 7.48 (t, J=7.0 Hz, 1H), 7.45-7.33(m, 1H), 7.16 (td, J=9.0, 3.5 Hz, 1H), 4.39 (s, 2H), 4.28 (s, 1H), 4.21(s, 1H), 3.85-3.72 (m, 2H), 3.71-3.48 (m, 9H), 3.39-3.32 (m, 2H), 3.17(s, 2H), 3.12 (s, 2H), 2.53-2.30 (m, 1H), 2.09-1.95 (m, 1H), 1.88 (ddd,J=13.8, 10.0, 6.3 Hz, 2H), 1.79-1.20 (m, 11H), 1.04 (dd, J=14.2, 8.7 Hz,2H), 0.90 (td, J=11.9, 11.3, 6.7 Hz, 1H), 0.79 (dd, J=13.2, 6.4 Hz, 4H),0.60 (d, J=11.1 Hz, 2H).

Example S-20. Preparation of4-(4-fluoro-3-(4-(2-(4-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-1H-1,2,3-triazol-1-yl)acetyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 1.107)

Step-1: Preparation of4-(3-(4-(2-bromoacetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To 4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (1.0 g, 2.48 mmol) in DCM (5 mL) was added Et₃N (3.58 mL,24.8 mmol) at RT and the mixture was slowly added to a previouslystirred solution of 2-bromoacetyl chloride (3.85 g, 24.8 mmol) in DCM(20 mL) at 0° C. The resultant mixture was stirred at RT for 10 min andmonitored by TLC and LC-MS. After completion, the reaction was quenchedwith saturated NaHCO₃ solution (30 mL) and extracted with DCM (200mL×2). The combined organic layers were washed with H₂O (100 mL), brine(50 mL), dried over anhydrous sodium sulphate and concentrated underreduced pressure to obtain a crude residue which was purified byCombiFlash chromatography to afford the title compound. LC-MS 487 [M+H]⁺

Step-2: Preparation of4-(3-(4-(2-azidoacetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To a stirred solution of4-(3-(4-(2-bromoacetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one (0.20 g, 0.411 mmol, 1 eq) in DMF (10 ml) was addedsodium azide (0.053, 0.83 m mol, 2 eq) at RT and the mixture was heatedat 80° C. for 2 h. The reaction was monitored by TLC. After completion,the mixture was diluted with EtOAc (100 mL). The organic layer waswashed with ice-cold water (80 mL×2), brine (50 mL), dried overanhydrous sodium sulphate and concentrated under reduced pressure toobtain a crude residue which was purified by CombiFlash chromatographyto afford the title compound. LC-MS 450 [M+H]⁺

Step-3: Preparation of4-(4-fluoro-3-(4-(2-(4-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-1H-1,2,3-triazol-1yl)acetyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of4-(3-(4-(2-azidoacetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one (0.120 g, 0.27 mmol, 1 eq) and(5S,8R,9S,10S,13S,14S,17R)-17-ethynyl-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(0.093 g, 0.32 mmol, 1.2 eq) in MeCN:H₂O (2:1) (6 mL) was addedCuSO₄.5H₂O (0.021 g, 0.13 mmol, 0.5 eq) and hydrazine hydrate (0.0085 g,0.26 mmol, 1 eq) and the resultant reaction mixture was heated at 80° C.for 16 h. The reaction was monitored by TLC and LC-MS. After completion,the mixture was diluted with water (30 mL) and extracted with EtOAc(40×2 mL). The combined organic layers were washed with brine (30 mL),dried with anhydrous sodium sulphate and concentrated under reducedpressure to obtain a crude which was purified by reversed phase HPLC toafford the title compound. LC-MS 764 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ12.60 (s, 1H), 8.30-8.23 (m, 1H), 8.01-7.79 (m, 3H), 7.68 (s, 1H),1.30-1.12 (m, 4H), 0.96 (s, 3H), 0.91 (s, 3H), 0.84 (d, J=10.5 Hz, 1H),0.48-0.38 (m, 2H). 7.48-7.33 (m, 2H), 7.25 (td, J=9.2, 5.9 Hz, 1H), 5.46(s, 1H), 5.40 (s, 1H), 5.05 (s, 1H), 4.34 (s, 2H), 3.71 (s, 1H),3.68-3.53 (m, 3H), 3.46 (s, 1H), 3.40 (s, 2H), 3.28 (d, J=7.5 Hz, 1H),3.20 (s, 1H), 2.42-2.21 (m, 2H), 2.10-1.99 (m, 1H), 1.95-1.78 (m, 3H),1.71 (dd, J=22.7, 10.8 Hz, 2H), 1.47-1.31 (m, 5H).

Example S-21. Preparation of4-(3-(4-(2-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)acetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one(Compound 1.110)

Step-1: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(10 g, 34.48 mmol) in benzene (330 mL) was added PTSA (1.54 g, 17.27mmol, 0.1 eq) followed by addition of ethylene glycol (9.63 g, 172.7mmol, 5.0 eq) at RT. The resultant mixture was heated at 150° C. 16 h.Reaction was monitored by TLC. Upon completion, the reaction mixture wasdiluted with water (200 mL) and extracted with EtOAc (350 mL). Theorganic layer was washed with saturated NaHCO₃ solution (100 mL), water(200 mL), brine (100 mL) dried over Na₂SO₄, filtered and concentratedunder reduced pressure to afford(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol.¹H NMR (400 MHz, DMSO-d₆) δ 4.40 (d, J=5.26 Hz, 1H), 3.81 (s, 3H),3.37-3.45 (m, 2H), 1.82 (brs, 2H), 1.70 (d, J=12.72 Hz, 2H), 1.59 (d,J=13.15 Hz, 2H), 1.52 (d, J=10.52 Hz, 3H), 1.35 (br. s., 1H), 1.23-1.33(m, 3H), 1.03-1.21 (m, 5H), 0.87 (dt, J=7.89, 19.95 Hz, 4H), 0.76 (s,3H), 0.61 (s, 3H)/

Step-2a: Preparation of 2-(2-bromoethoxy)tetrahydro-2H-pyran

To a stirred solution of 2-bromoethanol (10 g, 80.6 mmol) in diethylether (150 mL) was added PTSA (1.54 g, 8.0 mmol, 0.1 eq) followed byaddition of 3,4-dihydro-2H-pyran (8.83 g, 96.5 mmol, 1.2 eq) and theresultant mixture was stirred at RT for 16 h. Reaction was monitored byTLC. Upon completion, the reaction mixture was diluted with water (200mL) and extracted with EtOAc (350 mL). The organic layer was washed withsaturated NaHCO₃ solution (100 mL), water (200 mL), brine (100 mL) driedover Na₂SO₄, filtered and concentrated under reduced pressure to affordthe title compound as a yellowish liquid which was taken to next-stepwithout further purification. ¹H NMR (400 MHz, DMSO-d₆) δ 4.66 (t, J=3.3Hz, 1H), 3.93-3.85 (m, 1H), 3.82-3.66 (m, 2H), 3.66-3.57 (m, 2H),3.49-3.39 (m, 1H), 1.72 (d, J=9.2 Hz, 1H), 1.62 (d, J=2.2 Hz, 1H),1.53-1.40 (m, 4H).

Step-2: Preparation of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-(tetrahydro-2Hpyran-2-yloxy)ethoxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane]

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol(1 g, 2.99 mmol) in xylene (10 mL) was added NaNH₂ (50% suspension intoluene, 3 mL) and the mixture was heated at 150° C. for 1 h. Thereaction mixture was gradually cooled to RT, 2-(6-chlorohexyloxy)tetrahydro-2H-pyran (6 g, 29.9 mmol)) was added to it and the resultantmixture was again heated to 150° C. for 16 h. The reaction was monitoredby TLC. Upon completion, the mixture was cooled to RT, quenched slowlywith ice cold water (250 mL) and extracted with EtOAc (300 mL). Theorganic layer was washed with water (100 mL×2), brine (100 mL) driedover Na₂SO₄, filtered and concentrated under reduced pressure to obtaina crude product which was purified by CombiFlash chromatography toafford the title compound. LC-MS 463 [M+H]⁺

Step-3: Preparation of(5S,8R,9S,10S,13S,14S,17S)-17-(2-hydroxyethoxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-(tetrahydro-2H-pyran-2-yloxy)ethoxy)hexadecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane](0.635 g, 1.37 mmol) in THE (25 mL) water (5 mL) was added 6N HCl (15mL) at RT and the resultant reaction mixture was stirred at RT for 16 h.The reaction was monitored by TLC. Upon completion, the reaction mixturewas diluted with water (150 mL) and basified using with saturated NaHCO₃solution (pH ˜8). The aqueous layer was then extracted with EtOAc (200mL×3). The organic layer was washed with saturated NaHCO₃ solution (100mL), water (100 mL), brine (100 mL) dried over Na₂SO₄, filtered andconcentrated under reduced pressure to afford the title compound whichwas used in the next step without further purification. LC-MS 335 [M+H]⁺

Step-4: Preparation of2-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)acetic acid

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-(2-hydroxyethoxy)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(0.31 g, 0.928 mmol) in acetone (25 mL) was added Jones reagent (2.5 mL)at 0° C. dropwise over a period of 20 min. The resultant mixture wasstirred at 0° C. for 5 min. The reaction was monitored by TLC. Aftercompletion, water (10 mL) was added and the resulting precipitate wasfiltered over Buchner funnel. The product obtained was washed with water(5 mL×2) and n-pentane (5 mL×2), dried under vacuum to afford the titlecompound which was taken to next step without further purification.LC-MS 349 [M+H]⁺

Step-5: Preparation of4-(3-(4-(2-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)acetyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To a stirred solution of2-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)aceticacid (0.025 g, 0.071 mmol) in DMF (3 mL) was added HATU (0.032 g, 0.086mmol, 1.2 eq) at 0° C. and the resulting reaction mixture was stirredfor 10 min. DIPEA (0.062 mL, 0.358 mmol, 5 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.020 g, 0.0.5 mmol, 07 eq) were then successively addedto the mixture and the mixture was stirred at RT for 1 h. The reactionwas monitored by TLC & LC-MS. After completion, water (10 mL) was addedand the resulting precipitate was filtered over Buchner funnel. Theproduct obtained was washed with water (5 mL×2) and n-pentane (5 mL×2),dried under vacuum to obtain a crude product which was purified byreverse phase HPLC to afford the title compound. LC-MS 697 [M+H]⁺, ¹HNMR (400 MHz, MeOD-d₄) δ 8.41-8.34 (m, 1H), 8.00-7.92 (m, 1H), 7.86(dtd, J=18.0, 7.3, 1.4 Hz, 2H), 7.49 (t, J=6.3 Hz, 1H), 7.39 (t, J=6.0Hz, 1H), 7.17 (t, J=9.0 Hz, 1H), 4.61 (s, 2H), 4.39 (s, 2H), 4.21 (s,1H), 4.15 (s, 1H), 3.76 (d, J=7.6 Hz, 2H), 3.66 (q, J=5.9, 5.3 Hz, 2H),3.51 (s, 2H), 3.47-3.37 (m, 1H), 3.35 (d, J=4.2 Hz, 1H), 2.46 (d, J=15.1Hz, 1H), 2.37 (t, J=14.3 Hz, 1H), 2.22 (d, J=15.0 Hz, 1H), 2.11-1.80 (m,6H), 1.73 (d, J=13.8 Hz, 1H), 1.63 (d, J=10.8 Hz, 2H), 1.52 (m, 3H),1.39-1.27 (m, 6H), 1.25 (s, 1H), 1.06 (d, J=6.6 Hz, 3H), 0.99-0.88 (m,1H), 0.83 (s, 1H).

Example S-22. Preparation of4-(4-fluoro-3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 1.111) Step-1: Preparation of4-(3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To a stirred solution of6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid (0.10 g, 0.247 mmol) in DMF (15 mL) was added HATU (0.141 g, 0.371mmol, 1.5 eq) at 0° C. and the resulting reaction mixture was stirredfor 10 min. DIPEA (0.17 mL, 0.99 mmol, 4 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.119 g, 0.297 mmol, 0.1.2 eq) were then successivelyadded to the mixture and the mixture was stirred at RT for 4 h. Thereaction was monitored by TLC & LC-MS. After completion, water (10 mL)was added and the resulting precipitate was filtered over Buchnerfunnel. The product obtained was washed with water (25 mL×2) andn-pentane (25 mL×2), dried under vacuum to afford the title compound.LC-MS 753 [M+H]⁺

Step-6: Preparation of4-(4-fluoro-3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of4-(3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one(0.070 g, 0.093 mmol) in methanol (10 mL) was added NaBH₄ (0.0068 g,0.186 mmol, 2.0 eq) at 0° C. slowly and the mixture was stirred at RT 30min. The reaction was monitored by TLC. After completion, water (10 mL)was added and the resulting precipitate was filtered over Buchner funnelto obtain a crude residue which was purified by reversed phase HPLC toafford the title compound. LC-MS 755 [M+H]⁺, ¹H NMR (400 MHz, MeOD-d₄) δ8.37 (d, J=7.7 Hz, 1H), 7.95 (s, 1H), 7.86 (dt, J=15.5, 7.3 Hz, 2H),7.49 (d, J=7.1 Hz, 1H), 7.37 (d, J=6.1 Hz, 1H), 7.16 (t, J=9.0 Hz, 1H),4.39 (s, 2H), 3.82-3.61 (m, 4H), 3.49 (ddd, J=22.0, 12.8, 5.5 Hz, 6H),3.28 (d, J=7.6 Hz, 1H), 2.45 (t, J=7.6 Hz, 1H), 2.38 (t, J=7.6 Hz, 1H),2.00-1.91 (m, 1H), 1.85 (t, J=11.3 Hz, 1H), 1.78-1.47 (m, 10H),1.48-1.19 (m, 10H), 1.14 (d, J=14.1 Hz, 2H), 0.98 (q, J=10.6, 9.9 Hz,2H), 0.88 (s, 2H), 0.83 (s, 3H), 0.73 (d, J=8.8 Hz, 3H).

Example S-23. Preparation of4-(3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-3,3-difluoro-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one(Compound 1.114)

Step-1: Preparation of4-(3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To a stirred solution of6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid (0.50 g, 1.23 mmol) in DMF (20 mL) was added HATU (0.70 g, 1.84mmol, 1.5 eq) at 0° C. and the mixture was stirred for 10 min. DIPEA(1.1 mL, 6.18 mmol, 5 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.497 g, 1.23 mmol, 1.0 eq) were then successively addedto the reaction mixture at 0° C. and the resultant reaction mixture wasstirred at RT for 75 min. Reaction was monitored by TLC. Aftercompletion, water (10 mL) was added and the resulting precipitate wasfiltered over Buchner funnel. The product obtained was washed with water(10 mL×2) and n-pentane (10 mL×2), dried under reduced pressure toobtain a crude which was purified by CombiFlash chromatography to thetitle compound. LC-MS 753 [M+H]⁺

Step-2: Preparation of4-(3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-3,3-difluoro-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

To a stirred solution of 4-(3-(4-(6-((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one (0.25 g, 0.332 mmol) in 1,2-dichlorethane(10 mL) was added DAST (0.535 g, 3.32 mmol, 10 eq) dropwise and themixture was heated 50° C. for 1 h. The reaction was monitored by TLC.After completion, the mixture was quenched slowly with saturated NaHCO₃solution (50 mL) and extracted with DCM (100 mL×2). The combined organiclayers were washed with H₂O (100 mL), brine (50 mL), dried withanhydrous sodium sulphate and concentrated under reduced pressure toobtain a crude which was purified by reversed phase HPLC to afford thetitle compound. LC-MS 775 [M+H]⁺, ¹H NMR (400 MHz, DMSO-d₆) δ 12.59 (s,1H), 8.26 (d, J=7.8 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.89 (t, J=7.5 Hz,1H), 7.83 (t, J=7.5 Hz, 1H), 7.44 (t, J=6.7 Hz, 1H), 7.36 (t, J=6.0 Hz,1H), 7.23 (t, J=9.0 Hz, 1H), 4.33 (s, 2H), 3.62 (s, 1H), 3.59-3.47 (m,3H), 3.36 (d, J=9.2 Hz, 3H), 3.24 (q, J=7.5 Hz, 1H), 3.20-3.10 (m, 2H),2.33-2.22 (m, 2H), 1.87 (dd, J=16.6, 6.2 Hz, 2H), 1.81-1.56 (m, 6H),1.48 (q, J=9.3, 8.2 Hz, 6H), 1.39-1.01 (m, 12H), 0.98-0.81 (m, 2H), 0.79(s, 2H), 0.66 (d, J=6.7 Hz, 4H).

Example S-24. Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide(Compound 1.115)

Step-1: Preparation of(5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-01

To a stirred solution of(5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(1.0 g, 3.4 mmol) in acetic acid (20 mL)-H₂O (10 mL) was added zincmetal (10.0 g) at room temperature and the mixture was stirred at RT for16 h. The reaction was monitored by TLC. Upon completion, the mixturewas diluted with water (20 mL) and filtered through the celite bed, thecrude residue obtained from the celite bed was dissolved in methanol(100 mL) and concentrated under reduced pressure to afford the titlecompound. ¹HNMR (400 MHz, DMSO-d₆) δ 4.40 (d, J=4.8 Hz, 1H), 3.45-3.38(m, 1H), 1.81 (d, J=7.5 Hz, 1H), 1.71 (d, J=11.4 Hz, 1H), 1.66-1.55 (m,4H), 1.53-1.41 (m, 4H), 1.37-1.27 (m, 4H), 1.25-1.13 (m, 6H), 1.01 (br.s., 1H), 0.93 (dd, J=3.7, 12.9 Hz, 1H), 0.89-0.79 (m, 4H), 0.76 (s, 2H),0.61 (s, 2H).

Step-2a: Preparation of 2-(6-chlorohexyloxy)tetrahydro-2H pyran

To a stirred solution of 6-chlorohexan-1-ol (10 g, 73.5 mmol) in diethylether (100 mL) was added PTSA (1.4 g, 7.35 mmol, 0.1 eq) followed byaddition of 3,4-dihydro-2H-pyran (7.41 g, 88.0 mmol, 1.2 eq) at RT andthe mixture was stirred at RT for 16 h. The reaction was monitored byTLC. Upon completion, the mixture was diluted with water (200 mL) andextracted with EtOAc (350 mL). The organic layer was washed withsaturated NaHCO₃ solution (100 mL), water (200 mL), brine (100 mL) driedover Na₂SO₄, filtered and concentrated under reduced pressure to affordthe title compound which was taken to next step without furtherpurification. ¹HNMR (400 MHz, DMSO-d₆) δ 4.50 (br. s., 1H), 4.25 (t,J=6.80 Hz, 2H), 4.20 (s, 1H), 4.08-4.00 (m, 2H), 3.74-3.66 (m, 2H), 3.59(dd, J=6.36, 16.01 Hz, 2H), 2.66 (s, 2H), 1.90-1.81 (m, 2H), 1.67 (br.s., 1H), 1.45-1.38 (m, 2H), 1.27-1.20 (m, 2H), 1.17 (t, J=7.02 Hz, 2H).

Step-2: Preparation of2-(6-((5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexyloxy)tetrahydro-2H-pyran

To a stirred solution of(5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-ol(1.0 g, 2.40 mmol) in xylene (10 mL) was added NaNH₂ (50% suspension intoluene, 3 mL) and the mixture was heated at 150° C. for 1 h. Thereaction mixture was gradually cooled to RT,2-(6-chlorohexyloxy)tetrahydro-2H-pyran (4.2 g, 24.0 mmol)) was added toit and the resultant mixture was again heated to 150° C. for 16 h. Thereaction was monitored by TLC. Upon completion, the mixture was cooledto RT, quenched slowly with ice cold water (250 mL) and extracted withEtOAc (300 mL). The organic layer was washed with water (100 mL×2),brine (100 mL) dried over Na₂SO₄, filtered and concentrated underreduced pressure to obtain a crude residue which was purified byCombiFlash chromatography to afford the title compound. ¹HNMR (400 MHz,CDCl3) δ 5.82 (ddt, J=17.0, 10.3, 6.7 Hz, 1H), 5.05-4.91 (m, 2H), 4.57(d, J=4.4 Hz, 2H), 3.87 (ddd, J=11.2, 7.7, 3.1 Hz, 2H), 3.79-3.66 (m,2H), 3.55-3.45 (m, 2H), 3.24-3.32 (m, 2H), 3.26 (t, J=8.3 Hz, 1H),2.12-2.01 (m, 2H), 2.00-1.90 (m, 2H), 1.90-1.75 (m, 2H), 1.68-1.56 (m,8H), 1.54-1.41 (m, 8H), 1.41-1.31 (m, 4H), 1.26-1.17 (m, 4H), 0.94-0.8(m, 2H), 0.78 (s, 2H), 0.73 (s, 2H).

Step-3: Preparation of6-((5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-H1-cyclopenta[a]phenanthren-17-yloxy)hexan-1-ol

To a stirred solution of2-(6-((5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexyloxy)tetrahydro-2H-pyran(1.0 g, 2.17 mmol, 1 eq) in THE (28 mL)-H₂O (7 nit) was added 6N-HCl (20mL) at RT and the mixture was stirred at RT for 16 h. The reaction wasmonitored by TLC. Upon completion, the reaction mixture was diluted withwater (150 mL) and basified using with saturated NaHCO₃ solution (pH˜8). The aqueous layer was then extracted with EtOAc (200 mL×3). Theorganic layer was washed with saturated NaHCO₃ solution (100 mL), water(100 mL), brine (100 mL) dried over Na₂SO₄, filtered and concentratedunder reduced pressure to afford the title compound which was used inthe next step without further purification. ¹HNMR (400 MHz, DMSO-d₆) δ5.76 (s, 1H), 4.32 (t, J=5.3 Hz, 1H), 3.42-3.33 (m, 4H), 3.24 (t, J=8.3Hz, 1H), 2.01 (d, J=7.0 Hz, 1H), 1.95-1.86 (m, 1H), 1.77 (d, J=11.8 Hz,1H), 1.66-1.54 (m, 4H), 1.48-1.32 (m, 8H), 1.32-1.23 (m, 6H), 1.22-1.09(m, 6H), 1.01 (br. s., 1H), 0.90-0.79 (m, 4H), 0.75 (s, 2H), 0.69-0.61(m, 2H).

Step-4: Preparation of6-((5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid

To a stirred solution of6-((5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexan-1-ol(0.40 g, 1.09 mmol, 1 eq) in acetone (25 mL) was added Jones reagent(1.2 mL) at 0° C. dropwise over a period of 20 min. The resultantmixture was stirred at 0° C. for 5 min. The reaction was monitored byTLC. After completion. H₂O (20 mL) was added and the resultingprecipitate was filtered over Buchner funnel. The product obtained waswashed with water (5 mL×2) and n-pentane (5 mL×2), dried under vacuum toafford the title compound which was taken to next step without furtherpurification. ¹HNMR (400 MHz, DMSO-d₆) δ 11.96 (br. s., 1H), 3.28-3.18(m, 2H), 2.18 (t, J=7.5 Hz, 2H), 1.89 (d, J=5.7 Hz, 2H), 1.77 (d, J=11.4Hz, 2H), 1.53-1.40 (m, 8H), 1.39- 1.26 (m, 6H), 1.23 (br. s., 2H),1.23-1.06 (m, 10H), 0.96-0.80 (m, 4H), 0.75 (s, 2H), 0.71-0.56 (m, 2H).

Step-5: Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1 yl)-6-oxohexylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide

To a stirred solution of6-((5R,8R,9S,10S,13S,14S,17S)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid (0.190 g, 0.487 mmol, 1 eq) in DMF (12 mL) was added HATU (0.277 g,0.73 mmol, 1.2 eq) at 0° C. and the resulting reaction mixture wasstirred for 10 min. DIPEA (0.36 mL, 1.94 mmol, 4.0 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.156 g, 0.389 mmol, 0.8 eq) were then successively addedto the mixture and the mixture was stirred at RT for 2 h. The reactionwas monitored by TLC & LC-MS. After completion, water (10 mL) was addedand the resulting precipitate was filtered over Büchner funnel. Theproduct obtained was washed with water (5 mL×2) and n-pentane (5 mL×2),dried under vacuum to obtain a crude residue which was purified byreversed phase HPLC to afford the title compound. LC-MS 739[M+H]⁺. ¹HNMR400 MHz, DMSO-d₆) δ 12.59 (s, 1H), 8.26 (d, J=7.9 Hz, 1H), 8.01-7.93 (m,1H), 7.89 (t, J=7.2 Hz, 1H), 7.86-7.77 (m, 1H), 7.44 (br. s., 1H), 7.36(br. s., 1H), 7.23 (t, J=9.0 Hz, 1H), 4.33 (s, 2H), 3.62 (br. s., 1H),3.56 (br. s., 1H), 3.51 (br. s., 2H), 3.27-3.02 (m, 3H), 2.33 (br. s.,1H), 2.26 (br. s., 1H), 1.90 (br. s., 2H), 1.76 (br. s., 1H), 1.59-1.47(m, 8H), 1.35-1.28 (m, 6H), 1.20-1.16 (m, 10H), 1.13 (br. s., 2H), 0.83(br. s., 4H), 0.74 (br. s., 3H), 0.65 (d, J=6.6 Hz, 4H).

Example S-25. Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propioloyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 1.116)

Step-1: Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propioloyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred suspension of3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propiolicacid (0.50 g, 1.39 mmol) in DMF (20 mL) was added HATU (0.795 g, 2.095mmol, 1.5 eq) at 0° C. and the mixture was stirred for 10 min. DIPEA(1.45 mL, 8.35 mmol, 6 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.503 g, 1.25 mmol, 0.9 eq) were then successively addedto the reaction mixture at 0° C. and the resultant reaction mixture wasstirred at RT for 75 min. The reaction was monitored by TLC. Aftercompletion, water (10 mL) was added and the resulting precipitate wasfiltered over Buchner funnel. The product obtained was washed with water(10 mL×2) and n-pentane (10 mL×2), dried under reduced pressure toobtain a crude which was purified by CombiFlash chromatography to affordthe title compound. LC-MS 707 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 12.58(d, J=2.6 Hz, 1H), 8.24 (d, J=7.9 Hz, 1H), 8.00-7.92 (m, 1H), 7.87 (t,J=7.2 Hz, 1H), 7.83-7.73 (m, 1H), 7.41 (d, J=6.1 Hz, 1H), 7.33 (d, J=7.0Hz, 1H), 7.24-7.14 (m, 1H), 5.70-5.63 (m, 1H), 4.30 (d, J=7.9 Hz, 2H),3.74 (br. s., 1H), 3.63 (br. s., 3H), 3.39 (br. s., 1H), 3.24-3.10 (m,2H), 2.39-2.20 (m, 2H), 2.15-1.98 (m, 2H), 1.94-1.75 (m, 2H), 1.61 (br.s., 2H), 1.56 (br. s., 4H), 1.48-1.33 (m, 4H), 1.33-1.11 (m, 8H),0.97-0.91 (m, 3H), 0.76-0.73 (m, 3H).

Example S-26. Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17R)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 1.117)

Step-1: Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propioloyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred suspension of3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propiolicacid (0.50 g, 1.39 mmol) in DMF (20 mL) was added HATU (795 g, 2.095mmol, 1.5 eq) at 0° C. and the mixture was stirred for 10 min. DIPEA(1.45 mL, 8.35 mmol, 6 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.503 g, 1.25 mmol, 0.9 eq) were then successively addedto the reaction mixture at 0° C. and the resultant reaction mixture wasstirred at RT for 75 min. The reaction was monitored by TLC. Aftercompletion, water (10 mL) was added and the resulting precipitate wasfiltered over Buchner funnel. The product obtained was washed with water(10 mL×2) and n-pentane (10 mL×2), dried under reduced pressure toobtain a crude which was purified by CombiFlash chromatography to affordthe title compound. LC-MS 707 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 12.58(d, J=2.6 Hz, 1H), 8.24 (d, J=7.9 Hz, 1H), 8.00-7.92 (m, 1H), 7.87 (t,J=7.2 Hz, 1H), 7.83-7.73 (m, 1H), 7.41 (d, J=6.1 Hz, 1H), 7.33 (d, J=7.0Hz, 1H), 7.24-7.14 (m, 1H), 5.70-5.63 (m, 1H), 4.30 (d, J=7.9 Hz, 2H),3.74 (br. s., 1H), 3.63 (br. s., 3H), 3.39 (br. s., 1H), 3.24-3.10 (m,2H), 2.39-2.20 (m, 2H), 2.15-1.98 (m, 2H), 1.94-1.75 (m, 2H), 1.61 (br.s., 2H), 1.56 (br. s., 4H), 1.48-1.33 (m, 4H), 1.33-1.11 (m, 8H),0.97-0.91 (m, 3H), 0.76-0.73 (m, 3H).

Step-2: Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17R)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred suspension of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propioloyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(0.100 g, 0.141 mmol) in methanol (10 mL) was added 10% Pd/C (10 mg) andthe mixture was hydrogenated using hydrogen bladder for 16 h. Thereaction was monitored by TLC and LC-MS. After completion, the mixturewas filtered through celite bed and the filtrate was concentrated underreduced pressure to obtain a crude which was purified by reversed phaseHPLC to afford the title compound. LC-MS 711 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) 12.59 (s, 1H), 8.26 (d, J=7.8 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H),7.89 (t, J=7.6 Hz, 1H), 7.83 (t, J=7.5 Hz, 1H), 7.44 (dd, J=8.7, 5.0 Hz,1H), 7.35 (d, J=6.9 Hz, 1H), 7.24 (t, J=9.0 Hz, 1H), 4.33 (s, 2H), 3.98(dd, J=15.2, 8.0 Hz, 1H), 3.68-3.49 (m, 2H), 3.40-3.34 (m, 2H),3.23-3.11 (m, 2H), 3.05 (s, 1H), 3.00 (s, 1H), 2.44-2.34 (m, 1H), 2.28(d, J=14.2 Hz, 1H), 2.13-2.04 (m, 1H), 1.97-1.76 (m, 1H), 1.72-1.51 (m,6H), 1.45 (d, J=16.1 Hz, 6H), 1.36-1.13 (m, 8H), 0.98 (s, 2H), 0.91-0.58(m, 4H).

Example S-27. Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)prop-2-ynyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 1.118)

Step-1: Preparation of (5S,8R,9S,10S,13S,14S,17S)-17-(3-bromoprop-1ynyl)-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

To a stirred suspension of(5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-17-(3-hydroxyprop-1-ynyl)-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(1.0 g, 2.90 mmol, 1 eq) in DCM (20 mL) were successively addedtriphenyphosphine (2.28 g, 8.72 mmol, 3.0 eq) and carbon tetrabromide(1.98 gm, 5.98 mmol, 2 eq) at RT and the mixture was stirred at RT for 3h. The reaction was monitored by TLC. After completion, water (50 mL)was added and the aqueous layer was extracted with DCM (100 mL×2). Thecombined organic layers were washed with water (50 mL), brine (50 mL),dried over anhydrous sodium sulphate and concentrated under reducedpressure to afford crude residue which was purified by CombiFlashchromatography to afford the title compound. ¹H NMR (400 MHz, DMSO-d₆) δ5.36 (d, J=5.3 Hz, 1H), 4.49 (s, 2H), 4.30 (s, 2H), 2.29 (s, 2H), 1.90(br. s., 6H), 1.55 (br. s., 4H), 1.30 (br. s., 4H), 1.29-1.20 (m, 4H),0.98 (s, 2H), 0.74 (s, 4H).

Step-2: Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)prop-2ynyl) piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.20 g, 0.497 mmol) in EtOH (10 mL) was added DIPEA (0.44mL, 2.48 mmol, 5 eq) followed by the addition of(5S,8R,9S,10S,13S,14S,17S)-17-(3-bromoprop-1-ynyl)-17-hydroxy-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one(0.201 g, 0.497 mmol, 1.0 eq) and the mixture was heated at 80° C. for 1h. The reaction was monitored by TLC and LC-MS. After completion, themixture was concentrated under reduced pressure to obtain a cruderesidue which was purified by CombiFlash chromatography to afford thetitle compound. LC-MS 693 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₄) δ 12.58 (s,1H), 8.26 (d, 0.1=7.0 Hz, 1H), 7.98-7.91 (m, 1H), 7.90-7.76 (m, 2H),7.43 (br. s., 1H), 7.25 (d, J=4.8 Hz, 1H), 7.22-7.13 (m, 1H), 5.17 (s,1H), 4.32 (s, 2H), 3.63 (br. s., 2H), 3.35 (s, 2H), 3.17 (d, J=5.3 Hz,2H), 2.38 (br. s., 2H), 2.35-2.30 (m, 1H), 2.00 (d, J=14.5 Hz, 2H),1.89-1.75 (m, 4H), 1.60-1.52 (m, 6H), 1.47-1.40 (m, 6H), 1.22 (d, J=6.6Hz, 6H), 1.06-0.95 (m, 2H), 0.94 (s, 2H), 0.72 (s, 2H).

Example S-28. Preparation of(R)-N-(4-cyano-3-(trifluoromethyl)phenyl)-3-(4-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexylamino)phenylsulfonyl)-2-hydroxy-2-methylpropanamide(Compound 1.119)

Step-1: Preparation of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17R)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)propyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred suspension of4-(4-fluoro-3-(4-(3-((5S,8R,9S,10S,13S,14S,17S)-17-hydroxy-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)prop-2-ynyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(0.120 g, 0.173 mmol, 1 eq) in methanol (10 mL) was added 10% Pd/C (12mg) and the mixture was hydrogenated under hydrogen atmosphere for 16 h.The reaction was monitored by TLC and LC-MS. After completion, themixture was filtered through celite bed and the filtrate obtained wasconcentrated under reduced pressure to obtain a crude residue which waspurified by CombiFlash chromatography to afford the title compound.LC-MS 697 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 12.59 (s, 1H), 8.26 (d,J=7.5 Hz, 1H), 7.96 (d, J=7.9 Hz, 1H), 7.92-7.76 (m, 2H), 7.45-7.36 (m,1H), 7.30 (d, J=6.1 Hz, 1H), 7.22 (t, J=9.0 Hz, 1H), 4.32 (s, 2H), 3.99(br. s., 1H), 3.59 (br. s., 1H), 3.13 (br. s., 2H), 2.44-2.36 (m, 2H),2.33 (br. s., 2H), 2.25 (dd, J=11.6, 19.1 Hz, 4H), 2.09 (d, J=10.1 Hz,1H), 1.89 (t, J=13.6 Hz, 1H), 1.72 (br. s., 2H), 1.60 (br. s., 2H), 1.55(br. s., 2H), 1.50 (br. s., 2H), 1.46-1.42 (br. s., 4H), 1.37-1.33 (d,J=12.3 Hz, 4H), 1.31-1.04 (m, 8H), 0.98 (s, 2H), 0.80-0.74 (m, 2H).

Example S-29. Preparation of4-(4-fluoro-3-(4-(6-((8R,9S,13S,14S,17S)-3-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 2.2)

Step-1: Preparation of(8R,9S,13S,14S,175)-3-(4-methoxybenzyloxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-ol

To a stirred solution of(8R,9S,13S,14S,17S)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol(1.0 g, 3.6 mmol) in DMF (20 mL) was added sodium hydride (60%suspension in mineral oil; 0.22 g, 5.5 mmol) at RT and the mixture washeated at 50° C. for 2 h. The reaction was monitored by TLC. Uponcompletion, the mixture was quenched with ice-cold water (20 mL) andextracted with ethyl acetate (100 mL×2). The combined organic layerswere washed with chilled water (50 mL×3), brine (50 mL) dried withanhydrous sodium sulphate, concentrated under reduced pressure to obtaina crude which was purified by CombiFlash chromatography to afford thetitle compound. ¹HNMR (400 MHz, CDCl₃) δ 7.39-7.32 (m, J=8.8 Hz, 2H),7.26 (s, 1H), 7.20 (d, J=8.8 Hz, 1H), 6.95-6.85 (m, J=8.8 Hz, 2H), 6.77(dd, J=2.6, 8.3 Hz, 1H), 6.74-6.67 (m, 1H), 5.30 (s, 1H), 4.95 (s, 2H),3.82 (s, 3H), 3.73 (t, J=8.6 Hz, 2H), 3.54 (t, J=6.8 Hz, 1H), 2.84 (d,J=4.8 Hz, 2H), 2.31 (dd, J=3.1, 13.6 Hz, 1H), 2.20-2.10 (m, 2H),1.99-1.84 (m, 2H), 1.74-1.67 (m, 1H), 1.62 (d, J=14.0 Hz, 2H), 1.51-1.39(m, 3H), 1.24-1.14 (m, 1H), 0.89-0.78 (m, 2H).

Step-2a: Preparation of 2-(6-chlorohexyloxy)tetrahydro-2H pyran

To a stirred solution of 6-chlorohexan-1-ol (10 g, 73.5 mmol) in diethylether (100 mL) was added PTSA (1.4 g, 7.3.5 mmol, 0.1 eq) followed byaddition of 3,4-dihydro-2H-pyran (7.41 g, 88.0 mmol, 1.2 eq) at RT andthe mixture was stirred at RT for 16 h. The reaction was monitored byTLC. Upon completion, the mixture was diluted with water (200 mL) andextracted with EtOAc (350 mL). The organic layer was washed withsaturated NaHCO₃ solution (100 mL), water (200 mL), brine (100 mL) driedover Na₂SO₄, filtered and concentrated under reduced pressure to affordthe title compound (12.0 g, 74%) as a yellow liquid which was taken tonext step without further purification. ¹HNMR (400 MHz, DMSO-d₆) δ 4.50(br. s., 1H), 4.25 (t, J=6.80 Hz, 2H), 4.20 (s, 1H), 4.08-4.00 (m, 2H),3.74-3.66 (m, 2H), 3.59 (dd, J=6.36, 16.01 Hz, 2H), 2.66 (s, 2H),1.90-1.81 (m, 2H), 1.67 (br. s., 1H), 1.45-1.38 (m, 2H), 1.27-1.20 (m,2H), 1.17 (t, J=7.02 Hz, 2H).

Step-2: Preparation of2-(6-((8R,9S,13S,14S,17S)-3-(4-methoxybenzyloxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexyloxy)tetrahydro-2H-pyran

To a stirred solution of(8R,9S,13S,14S,17S)-3-(4-methoxybenzyloxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-ol(0.50 g, 1.27 mmol) in xylene (5 mL) was added NaNH₂ (50% suspension intoluene, 1.5 mL) and the mixture was heated at 150° C. for 1 h. Thereaction mixture was gradually cooled to RT,2-(6-chlorohexyloxy)tetrahydro-2H-pyran (2.8 g, 12.5 mmol, 10 eq) wasadded to it and the resultant mixture was again heated to 150° C. for 16h. The reaction was monitored by TLC. Upon completion, the mixture wascooled to RT, quenched slowly with ice cold water (250 mL) and extractedwith EtOAc (300 mL). The organic layer was washed with water (100 mL×2),brine (100 mL) dried over Na₂SO₄, filtered and concentrated underreduced pressure to obtain a crude residue which was purified byCombiFlash chromatography to afford the title compound. ¹HNMR (400 MHz,DMSO-d₆) δ 7.37-7.31 (m, J=8.3 Hz, 2H), 7.14 (d, J=8.3 Hz, 1H),6.94-6.90 (m, J=8.3 Hz, 2H), 6.72 (d, J=7.9 Hz, 1H), 6.67 (br. s., 1H),4.53 (br. s., 2H), 3.76-3.73 (m, 3H), 3.64-3.56 (m, 2H), 3.45-3.36 (m,4H), 2.75 (br. s., 2H), 2.26 (br. s., 2H), 2.10 (d, J=12.7 Hz, 2H), 1.99(s, 2H), 1.90 (d, J=8.3 Hz, 2H), 1.80 (br. s., 2H), 1.69 (br. s., 2H),1.60 (br. s., 2H), 1.55-1.39 (m, 8H), 1.35-1.27 (m, 6H), 1.25 (br. s.,2H), 0.90-0.81 (m, 2H).

Step-3: Preparation of6-((8R,9S,13S,14S,17S)-3-(4-methoxybenzyloxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexan-1-ol

To a stirred solution of2-(6-((8R,9S,13S,14S,17S)-3-(4-methoxybenzyloxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexyloxy)tetrahydro-2H-pyran(0.42 g, 0.72 mmol) in THE (12 mL) water (3 mL) was added 6-N HCl (9 mL)at RT and the resultant mixture was stirred at RT for 16 h. The reactionwas monitored by TLC. Upon completion, the reaction mixture was dilutedwith water (40 mL) and basified using with saturated NaHCO₃ solution(pH˜8). The aqueous layer was then extracted with EtOAc (50 mL×3). Theorganic layer was washed with saturated NaHCO₃ solution (40 mL), water(40 mL), brine (50 mL) dried over Na₂SO₄, filtered and concentratedunder reduced pressure to afford the title compound. LC-MS 493[M+H]⁺

Step-4: Preparation of6-((8R,9S,13S,14S,17S)-3-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid

To a stirred solution of6-((8R,9S,13S,14S,17S)-3-(4-methoxybenzyloxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexan-1-ol(0.18 g, 0.365 mmol) in acetone (18 mL) was added Jones reagent (1.2 mL)at 0° C. dropwise over a period of 20 min. The resultant mixture wasstirred at 0° C. for 5 min. The reaction was monitored by TLC. Aftercompletion, water (20 mL) was added and the resulting precipitate wasfiltered over Büchner funnel. The product obtained was washed with water(5 mL×2) and n-pentane (5 mL×2), dried under vacuum to afford the titlecompound which was taken to next step without further purification.LC-MS 387[M+H]⁺

Step-5: Preparation of4-(4-fluoro-3-(4-(6-((8R,9S,13S,14S,17S)-3-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of6-((8R,9S,13S,14S,17S)-3-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yloxy)hexanoicacid (0.190 g, 0.466 mmol) in DMF (8 mL) was added HATU (0.277 g, 0.699mmol, 1.5 eq) at 0° C. and the resulting reaction mixture was stirredfor 10 min. DIPEA (0.24 mL, 1.86 mmol, 4.0 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.149 g, 0.373 mmol, 0.8 eq) were then successively addedto the mixture and the mixture was stirred at RT for 2 h. The reactionwas monitored by TLC & LC-MS. After completion, water (10 mL) was addedand the resulting precipitate was filtered over Buchner funnel. Theproduct obtained was washed with water (5 mL×2) and n-pentane (5 mL×2),dried under vacuum to obtain a crude residue which was purified byreversed phase HPLC to afford the title compound. LC-MS 735[M+H]⁺. ¹HNMR(400 MHz, DMSO-d₆) δ 12.59 (s, 1H), 8.99 (s, 1H), 8.26 (d, J=7.8 Hz,1H), 7.96 (d, J=7.9 Hz, 1H), 7.89 (t, J=7.6 Hz, 1H), 7.83 (t, J=7.5 Hz,1H), 7.48-7.32 (m, 2H), 7.23 (t, J=9.0 Hz, 1H), 7.06-6.98 (m, 1H), 6.49(dd, J=8.3, 2.6 Hz, 1H), 6.42 (d, J=2.6 Hz, 1H), 4.32 (s, 2H), 3.63 (d,J=6.0 Hz, 1H), 3.54 (d, J=22.1 Hz, 3H), 3.39 (dt, J=17.2, 5.3 Hz, 4H),3.15 (dd, J=13.2, 8.4 Hz, 2H), 2.70 (d, J=10.9 Hz, 2H), 2.35 (d, J=7.7Hz, 1H), 2.28 (d, J=7.6 Hz, 1H), 2.24-2.16 (m, 1H), 2.12-2.03 (m, 2H),2.02-1.81 (m, 3H), 1.75 (t, J=7.5 Hz, 1H), 1.64-1.41 (m, 4H), 1.41-1.10(m, 8H), 0.70 (d, J=6.7 Hz, 3H).

Example S-30. Preparation of4-(4-fluoro-3-(4-(6-((2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)(methyl)amino)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 2.4)

Step-1: Preparation of 6-(methylamino)hexanoic acid hydrochloride

To N-methylcaprolactam (5 g, 39.3 mmol) was added conc. HCl (25 mL)-H₂O(36 mL) at RT and the mixture was refluxed at 110° C. for 16 h. Thereaction was monitored by TLC. After 16 h, the mixture was concentratedunder reduced pressure. To the residue obtained was added 10 mL of H₂Oand again concentrated. The crude obtained was triturated withacetone/n-pentane (1:5) twice to afford the title compound. ¹H NMR (400MHz, DMSO-d₆) δ 12.05 (1H, s, br), 8.70 (2H, br), 3.6 (s, 3H), 2.18-2.23(2H, m), 2.77-2.86 (2H, m), 1.44-1.62 (2H, m), 1.24-1.34 (2H, m).

Step-2: Preparation of 6-(tert-butoxycarbonyl(methyl)amino)hexanoic acid

To a stirred solution 6-(methylamino)hexanoic acid hydrochloride (1.5 g,10.3 mmol) in 1,4-dioxane was added 1N-NaOH (20.7 mL, 2.5 eq) followedby the addition of di-tert-butyl dicarbonate (2.7 g, 12.4 mmol, 1.2 eq)at 0° C. and the mixture was stirred at RT for 16 h. The reaction wasmonitored by TLC. Upon completion, the mixture was concentrated underreduced pressure to afford the title compound which was taken forwardwithout further purification. ¹H NMR (400 MHz, DMDO-d₆) δ 11.97 (br s,1H), 3.57 (s, 3H), 3.12 (t, J=6.6 Hz, 2H), 2.29 (t, J=7.1 Hz, 2H), 2.19(t, J=7.1 Hz, 2H), 1.52 (d, J=6.4 Hz, 2H), 1.38 (s, 9H), 1.20 (br s,2H).

Step-3: Preparation of tert-butyl6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1 yl)-6-oxohexyl(methyl)carbamate

To a stirred suspension of 6-(tert-butoxycarbonyl(methyl)amino)hexanoicacid (0.20 g, 0.8 mmol) in DMF (4 mL) was added HATU (0.62 g, 1.6 mmol,2 eq) at 0° C. and the mixture was stirred at 0° C. for 10 min. DIPEA(0.45 mL, 2.4 mmol, 3 eq) and4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.36 g, 0.89 mmol, 1.1 eq) were then successively addedto the reaction mixture at 0° C. and the resultant reaction mixture wasstirred at RT for 16 h. The reaction was monitored by TLC. Aftercompletion, the mixture was diluted with H₂O (50 mL) and extracted withEtOAc (50 mL×2). The combined organic layers were washed with water (50mL), brine (50 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to obtain a crude which was purifiedby CombiFlash chromatography to afford the title compound. LC-MS 594[M+H]⁺.

Step-4: Preparation of4-(4-fluoro-3-(4-(6-(methylamino)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To tert-butyl6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexyl(methyl)carbamate(0.23 g, 0.38 mmol) in DCM (5 mL) was added TFA (0.3 mL) at 0° C. slowlyand the mixture was heated at 60° C. for 2 h. The reaction was monitoredby TLC and LC-MS. Upon completion, the mixture was concentrated underreduced pressure to obtain a crude residue which was triturated withn-pentane (10 mL×2) to afford the title compound as a trifluoroacetatesalt. LC-MS 494 [M+H]⁺.

Step 5a: Preparation of(4-fluorophenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone

A mixture of 6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene (8.5 g,31.44 mmol) in DCM (100 mL) was cooled to 0° C., and then4-fluorobenzoyl chloride (5.73 g, 36.16 mmol) and AlCl₃ (4.61 g, 34.58mmol) was added. The reaction mixture was stirred at rt for 4 h. TLCshowed the reaction mixture was complete. The solution was poured intowater and extracted with DCM/MeOH (100 mL×3). The organic layer waswashed with brine, dried over NaSO₄, and concentrated in vacuo. Thecrude product was purified by column chromatography to afford the titlecompound.

Step 5b: Preparation of(4-(2-hydroxyethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3yl)methanone

A mixture of ethylene glycol (25.59 g, 412.8 mmol) in DMF (100 mL) andcooled to 0° C., NaH (3.3 g, 82.56 mmol) was added slowly. The reactionmixture was stirred at rt for 1 h. A solution of(4-fluorophenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone(10.8 g, 27.52 mmol) in DMF (20 mL) was added dropwise at 0° C. Thereaction mixture was stirred at 90° C. for 1 h. TLC showed the mixturewas complete. The solution was concentrated in vacuo, and purified bycolumn chromatography to afford the title compound. LC-MS 435.3 [M+1]⁺.¹H NMR (400 MHz, DMSO-d₆) δ 7.70-7.65 (m, 3H), 7.34-7.31 (m, 3H), 7.0(dd, J=6.8, 2.4 Hz, 1H), 6.95-6.88 (m, 4H), 4.88 (t, J=1.6 Hz, 1H), 4.01(t, J=4.8 Hz, 2H), 3.84 (s, 3H), 3.72 (s, 3H), 3.71-3.67 (m, 2H).

Step-5c: Preparation of(4-(2-bromoethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone

To a stirred solution of(4-(2-hydroxyethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone(200 mg, 0.460 mmol) in DMF (5 mL) was added PBr₃ (0.87 mL, 0.92 mmol, 2eq) at 0° C. and the mixture was stirred at 50° C. for 1 h. The reactionwas monitored by TLC. Upon completion, the mixture was quenched withsaturated NaHCO₃ solution and extracted with EtOAc (60 mL×3). Thecombined organic layers were washed with water (60 mL), brine (60 mL)and dried over Na₂SO₄, filtered and concentrated under reduced pressureto afford the title compound. LC-MS 497 [M+H]⁺.

Step-5: Preparation of4-(4-fluoro-3-(4-(6-((2-(4-(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)(methyl)amino)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of4-(4-fluoro-3-(4-(6-(methylamino)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one trifluoroacetate (0.080 g, 0.162 mmol) in EtOH (2mL) was added DIPEA (0.15 mL, 0.81 mmol, 5 eq) followed by the additionof (4-(2-bromoethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone (0.08 g, 0.162 mmol, 1 eq) andthe mixture was heated at 80° C. for 16 h. The reaction was monitored byTLC and LC-MS. Upon completion, the mixture was concentrated underreduced pressure to obtain a crude which was purified by CombiFlashchromatography to afford the title compound. LC-MS 910 [M+H]⁺.

Step-6: Preparation of4-(4-fluoro-3-(4-(6-((2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)(methyl)amino)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one (Compound 2.4)

To a stirred solution of4-(4-fluoro-3-(4-(6-((2-(4-(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)(methyl)amino)hexanoyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one (100 mg, 0.11 mmol) in DCM (2 mL) was added BBr₃(1M in DCM; 1.09 mL, 1.1 mmol, 10 eq) at 0° C. and the mixture wasstirred at RT for 16 h. The reaction was monitored by LC-MS and TLC.Upon completion, the mixture was quenched with saturated NaHCO₃ solution(60 mL). The aqueous layer was then extracted with EtOAc (60 mL×3). Thecombined organic layers were washed with water (60 mL), brine (60 mL)and dried over Na₂SO₄, filtered and concentrated under reduced pressureto obtain a crude which was purified by reversed-phase HPLC to affordCompound 2.4. LC-MS 882 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD-d₄) δ 8.54 (s,1H), 8.39-8.31 (m, 1H), 7.98-7.91 (m, 1H), 7.83 (p, J=7.3 Hz, 1H),7.75-7.66 (m, 2H), 7.47 (d, J=8.6 Hz, 2H), 7.42 (d, J=8.7 Hz, 1H), 7.36(s, 1H), 7.25 (d, J=2.3 Hz, 1H), 7.17 (dd, J=8.6, 6.7 Hz, 2H), 6.91-6.81(m, 3H), 6.62 (d, J=8.3 Hz, 2H), 4.38 (s, 2H), 4.16 (d, J=4.9 Hz, 2H),3.78-3.69 (m, 2H), 3.63 (d, J=7.0 Hz, 2H), 3.35 (s, 2H), 2.93 (s, 1H),2.59 (s, 1H), 2.42 (d, J=6.8 Hz, 3H), 1.58 (d, J=15.1 Hz, 4H).

Example S-31. Preparation of4-(4-fluoro-3-(4-(2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(Compound 2.13)

Step-1a: Preparation of(4-(2-bromoethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone

To a stirred solution of(4-(2-hydroxyethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone(200 mg, 0.460 mmol) in DMF (5 mL) was added PBr₃ (0.87 mL, 0.92 mmol, 2eq) at 0° C. and the mixture was stirred at 50° C. for 1 h. The reactionwas monitored by TLC. Upon completion, the mixture was quenched withsaturated NaHCO₃ solution and extracted with EtOAc (60 mL×3). Thecombined organic layers were washed with water (60 mL), brine (60 mL)and dried over Na₂SO₄, filtered and concentrated under reduced pressureto afford(4-(2-bromoethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone.LC-MS 497 [M+H]⁺

Step-1: Preparation of4-(4-fluoro-3-(4-(2-(4-(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred suspension of4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (150 mg, 0.372 mmol) in DMF (5 mL) were successively addedK2CO₃ (102 mg, 0.744 mmol, 2 eq), NaI (11.16 mg, 0.0743 mmol, 0.2 eq)and(4-(2-bromoethoxy)phenyl)(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophen-3-yl)methanone(222 mg, 0.444 mmol, 1.2 eq) and the resultant mixture was stirred at100° C. for 6 h. The reaction was monitored by TLC. After completion,the mixture was diluted with water (20 mL). The resulting precipitatewas filtered over Buchner funnel to obtain a crude product which waspurified by combi flash chromatography to afford4-(4-fluoro-3-(4-(2-(4-(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one.LC-MS 783[M+H]⁺

Step-2: Preparation of4-(4-fluoro-3-(4-(2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To a stirred solution of4-(4-fluoro-3-(4-(2-(4-(6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one(100 mg, 0.127 mmol) in DCM (5 mL) was added BBr₃ (1M in DCM; 1.5 mL,1.53 mmol, 12 eq) at 0° C. and the mixture was stirred at RT for 16 h.The reaction was monitored by LC-MS and TLC. Upon completion, themixture was quenched with saturated NaHCO₃ solution (60 mL). The aqueouslayer was then extracted with EtOAc (60 mL×3). The combined organiclayers were washed with water (60 mL), brine (60 mL) and dried overNa₂SO₄, filtered and concentrated under reduced pressure to obtain acrude which was purified by reversed phase HPLC to afford4-(4-fluoro-3-(4-(2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one.LC-MS 755 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 12.60 (s, 1H), 9.79 (s,1H), 9.74 (s, 1H), 8.25 (d, J=7.9 Hz, 1H), 7.96 (d, J=8.1 Hz, 1H), 7.88(t, J=7.7 Hz, 1H), 7.81 (t, J=7.5 Hz, 1H), 7.65 (d, J=8.5 Hz, 2H), 7.41(t, J=6.9 Hz, 1H), 7.34 (d, J=2.3 Hz, 1H), 7.31 (dd, J=6.7, 2.1 Hz, 1H),7.27-7.19 (m, 2H), 7.19-7.14 (m, 2H), 6.92 (d, J=8.7 Hz, 2H), 6.85 (dd,J=8.6, 2.3 Hz, 1H), 6.70-6.64 (m, 2H), 4.32 (s, 2H), 4.10 (dt, J=7.6,5.3 Hz, 2H), 3.59 (s, 2H), 3.17 (d, J=5.2 Hz, 1H), 3.13 (s, 2H), 2.69(t, J=5.5 Hz, 2H), 2.44 (s, 2H), 2.34 (s, 1H).

Example S-32. Synthesis of4-(4-fluoro-3-(4-(2-(4-(1-(4-hydroxyphenyl)-2-phenylbut-1-enyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)pthalazin-1(2H)-one(Compound 2.14) Step-1: Preparation of4-(1-(4-(2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1yl)methyl)benzoyl)piperazin-1 yl)ethoxy)phenyl)-2phenylbut-1-enyl)phenyl pivalate

To a stirred solution of4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-onehydrochloride (0.10 g, 0.197 mmol) in EtOH (2 mL) was added DIPEA (0.91mL, 4.97 mmol, 5 eq) followed by the addition of4-(1-(4-(2-bromoethoxy)phenyl)-2-phenylbut-1-enyl)phenyl pivalate (0.119g, 0.296 mmol, 1.5 eq) and the mixture was heated at 80° C. for 16 h.The reaction was monitored by TLC and LC-MS. Upon completion, themixture was concentrated under reduced pressure to obtain a crude whichwas purified by reversed-phase HPLC to afford the title compound. LC-MS793 [M+H]⁺.

Step-2: Preparation of 4-(4-fluoro-3-(4-(2-(4-(1-(4-hydroxyphenyl)-2phenylbut-1-enyl)phenoxy)ethyl)piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one

To4-(1-(4-(2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)ethoxy)phenyl)-2-phenylbut-1-enyl)phenylpivalate (0.035 g, 0.044 col) was added 1.25 N HCl in EtOH (2 mL) andthe mixture was stirred at RT for 24 h. The reaction was monitored byTLC and LC-MS. After completion, the mixture was concentrated underreduced pressure to obtain a crude which was purified by reversed phaseHPLC to afford Compound 2.14. LC-MS 709 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 12.59 (br s, 1H), 9.41 (s, 1H), 9.16 (s, 1H), 8.21-8.29 (m,1H), 7.94-8.00 (m, 1H), 7.88 (d, J=6.1 Hz, 1H), 7.81 (d, J=4.8 Hz, 1H),7.41 (br s, 1H), 7.32 (br s, 1H), 7.12-7.25 (m, 2H), 7.08 (d, J=8.3 Hz,2H), 6.93 (d, J=8.8 Hz, 3H), 6.66-6.79 (m, 2H), 6.59 (dd, J=8.8, 3.5 Hz,2H), 6.39 (d, J=8.8 Hz, 1H), 4.32 (d, J=4.4 Hz, 2H), 4.08 (d, J=5.7 Hz,1H), 3.92 (d, J=5.3 Hz, 1H), 3.60 (d, J=18.0 Hz, 2H), 3.14 (d, J=19.7Hz, 2H), 2.73 (d, J=5.7 Hz, 1H), 2.63 (br s, 1H), 2.39 (t, J=7.2 Hz,4H), 0.84 (t, J=7.5 Hz, 3H).

Example S-33. Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-N-(6-(3-(5-(8-ethyl-7-oxo-5,6,7,8-tetrahydropyrazino[2,3-b]pyrazin-2-yl)-6-methylpyridin-2-yl)-1H-1,2,4-triazol-1-yl)hexyl)-2-fluorobenzamide(Compound 3.1)

Step-1: Preparation of 3-bromo-2-methyl-6-(1H-1,2,4-triazol-3yl)pyridine

To3-bromo-2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)pyridinic(1.6 g, 4.95 mmol) was added 2M-HCl in MeOH (15 mL) and resultingmixture was stirred at RT for 16 h. The reaction was monitored by TLC &LC-MS. After completion, the mixture was concentrated under reducedpressure to afford the title compound. LC-MS 239 [M+H]⁺

Step-2a: Preparation of 2-(6-iodohexyl)isoindoline-1,3-dione

To a solution of potassium 1,3-dioxoisoindolin-2-ide (5.0 g, 26.99 mmol)in DMF (50 mL) was added 1,6-Diiodohexane (22.8 g, 67.49 mmol) and themixture was heated at 70° C. for 2.5 h. The reaction was monitored byTLC. After completion, the mixture was diluted with H₂O (100 mL) andextracted with EtOAc (200 mL×2). The combined organic layers were washedwith brine (100 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to obtain a crude which was purifiedby CombiFlash chromatography to afford the title compound. LC-MS 358[M+H]⁺

Step-2: Preparation of 2-(6-(3-(5-bromo-6-methylpyridin-2yl)-1H-1,2,4-triazol-1 yl)hexyl)isoindoline-1,3-dione

To a solution of 3-bromo-2-methyl-6-(1H-1,2,4-triazol-3-yl)pyridine (1.4g, 5.08 mmol) in DMF (25 mL) was added K2CO₃ (1.75 g, 12.70 mmol) at 0°C. and the mixture was stirred at RT for 15 min.2-(6-iodohexyl)isoindoline-1,3-dione (1.99 g, 5.58 mmol) and then addedto the mixture and the resultant mixture was heated at 70° C. for 1.5 h.The reaction was monitored by TLC. Upon completion, the mixture wasdiluted with H₂O (50 mL) and extracted with EtOAc (100 mL×2). Thecombined organic layers were washed with brine (100 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toobtain a crude which was purified by CombiFlash chromatography to affordthe title compound. LC-MS 468[M+H]⁺

Step 3a: Preparation of1-ethyl-7-(trimethylstannyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one

To a stirred solution of7-bromo-1-ethyl-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one (0.5 g, 1.94mmol) in DMF (8 mL) was added hexamethylditin (0.48 mg, 2.33 mmol, 1.2eq) at RT and the mixture was degassed under nitrogen for 15 min.Pd(PPh₃)₄ (0.224 mg, 0.194 mmol, 0.1 eq) was then added to the mixtureresultant reaction mixture was heated at 100° C. for 90 min. Thereaction monitored by TLC. After completion, the mixture was dilutedwith water (200 mL) and extracted with EtOAC (200 mL×2). The combinedorganic layers were washed with water (100 mL), brine (150 mL) driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto afford a crude which was purified by CombiFlash chromatography toafford the title compound. LC-MS 343 [M+H]⁺

Step-3: Preparation of2-(6-(3-(5-(8-ethyl-7-oxo-5,6,7,8-tetrahydropyrazino[2,3-b]pyrazin-2yl)-6-methylpyridin-2-yl)-1H-1,2,4-triazol-1yl)hexyl)isoindoline-1,3-dione

To a stirred solution of2-(6-(3-(5-bromo-6-methylpyridin-2-yl)-1H-1,2,4-triazol-1-yl)hexyl)isoindoline-1,3-dione (0.5 g, 1.94 mmol) in DMF (8 mL) were added1-ethyl-7-(trimethylstannyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one(0.507 g, 1.48 mmol, 1.7 eq) and triethylamine (0.48 mL, 3.50 mmol, 4.0eq) at RT and the mixture was degassed under nitrogen for 15 min.Tri(o-tolyl)phosphine (0.053 g, 0.0175 mmol, 0.2 eq) and Pd₂(dba)₃(0.080 mg, 0.087 mmol, 0.1 eq) were then successively added to themixture and the resultant mixture was heated at 100° C. for 16 h. Thereaction was monitored by TLC. After completion, the mixture was dilutedwith water (50 mL) and extracted with EtOAc (200 mL×2). The combinedorganic layers was washed with brine (100 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford acrude which was purified by CombiFlash chromatography to afford thetitle compound. LC-MS 566 [M+H]⁺

Step 4: Preparation of5-(5-amino-2-(p-tolyloxy)phenyl)-1-methyl-3-(methylamino)pyridin-2(1H)-one

To a stirred solution of2-(6-(3-(5-(8-ethyl-7-oxo-5,6,7,8-tetrahydropyrazino[2,3-b]pyrazin-2-yl)-6-methylpyridin-2-yl)-1H-1,2,4-triazol-1-yl)hexyl)isoindoline-1,3-dione(0.20 g, 0.353 mmol) in EtOH (10 mL) was added NH₂NH₂.H₂O (0.52 mL, 1.06mmol, 3.0 eq) and the mixture was heated at 100° C. for 4 h. Thereaction monitored by TLC and LC-MS. After completion, the mixture wasdiluted with water (200 mL) and extracted with 10% MeOH/DCM (200 mL×3).The combined organic layers were washed with water (100 mL), brine (150mL) dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford the title compound. LC-MS 436 [M+H]⁺

Step-5: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-N-(6-(3-(5-(8-ethyl-7-oxo-5,6,7,8-tetrahydropyrazino[2,3-b]pyrazin-2-yl)-6-methylpyridin-2-yl)-1H-1,2,4-triazol-1yl)hexyl)-2-fluorobenzamide

To a stirred solution of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoicacid (0.13 g, 0.287 mmol) in DMA (5 mL) was added HBTU (0.130 g, 0.345mmol, 1.2 eq) at 0° C. and the resulting mixture was stirred for 10 min.DIPEA (0.150 mL, 0.575 mmol, 2.0 eq) and5-(5-amino-2-(p-tolyloxy)phenyl)-1-methyl-3-(methylamino)pyridin-2(1H)-one(0.150 g, 0.345 mmol, 1.2 eq) were then successively added to themixture and the resultant mixture was stirred at RT for 2 h. Thereaction was monitored by TLC & LC-MS. After completion, water (20 mL)was added and the resulting precipitate was filtered over Buchnerfunnel. The product obtained was washed with water (5 mL×2) andn-pentane (5 mL×2), dried under vacuum to obtain a crude residue whichwas purified by reversed phase HPLC to afford the title compound. LC-MS869 [M+H]⁺, ¹HNMR (400 MHz, MeOD-d₄) δ 8.17 (dd, J=5.2, 3.1 Hz, 2H),8.06-7.95 (m, 4H), 7.86 (s, 1H), 7.80 (t, J=8.2 Hz, 1H), 7.41-7.29 (m,2H), 4.27 (s, 2H), 4.18 (q, J=7.0 Hz, 2H), 3.38 (t, J=6.9 Hz, 2H), 2.76(s, 3H), 2.06-1.91 (m, 3H), 1.61 (d, J=8.6 Hz, 3H), 1.58 (s, 6H),1.50-1.43 (m, 3H), 1.26 (m, 2H).

Example S-34. Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-N-(6-(4-ethyl-6-(2-methyl-6-(1H-1,2,4-triazol-3-yl)pyridin-3-yl)-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)-2-fluorobenzamide(Compound 3.2)

Step-1: Preparation of ethyl 2-(3,5-dibromopyrazin-2 ylamino)acetate

To a stirred solution of 3,5-dibromopyrazin-2-amine (20.0 g, 79.6 mmol)in DMF (100 mL) was added Cs₂CO₃ (36.2 g, 102.8 mmol) at 0° C. and themixture was stirred for 15 min. Ethyl 2-bromoacetate (10.4 g, 94.8 mmol)was then added to the mixture and the mixture was heated at 80° C. for 3h. The reaction was monitored by TLC. Upon completion, the mixture wasdiluted with water (200 mL) to obtain a precipitate which was filteredover Buchner funnel and dried under vacuum to afford the title compound.LC-MS 338 [M+H]⁺

Step-2: Preparation of ethyl 2-(5-bromo-3-(ethylamino)pyrazin-2ylamino)acetate

To a stirred solution of ethyl 2-(3,5-dibromopyrazin-2-ylamino)acetate(1.1 g, 3.22 mmol) in NMP (9 mL) was added DIPEA (2.82 mL, 16.2 mmol, 5eq) and ethanamine hydrochloride (70% solution in water; 1.04 mL, 12.9mmol, 2.0 eq) and the mixture was heated at 120° C. in sealed tube for16 h. The reaction was monitored by TLC and LC-MS. After completion. Themixture was diluted with NaHCO₃ solution (100 mL) and extracted withEtOAc (200 mL×2). The combined organic layers were washed with brine(100 mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to obtain a get crude which was purified by CombiFlashchromatography to afford the title compound LC-MS 303[M+H]⁺

Step-3: Preparation of7-bromo-1-ethyl-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one

To a stirred solution of ethyl2-(5-bromo-3-(ethylamino)pyrazin-2-ylamino)acetate (0.97 g, 3.20 mmol)in methanol (1 mL) was added AcOH (5 mL) at RT and the mixture washeated 120° C. in sealed tube for 16 h. The reaction was monitored byTLC and LC-MS. After completion, the mixture was diluted with saturatedNaHCO₃ solution (100 mL) and extracted with EtOAc (100 mL×3). Thecombined organic layers were washed with brine (50 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toobtain a crude which q was purified by CombiFlash chromatography toafford the title compound. LC-MS 257[M+H]⁺

Step-4: Preparation of2-(6-(6-bromo-4-ethyl-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)isoindoline-1,3-dione

To a stirred solution of7-bromo-1-ethyl-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one (0.8 g, 3.11mmol) in DMF (25 mL) was added Cs₂CO₃ (2.02 g, 6.24 mmol, 2 eq) at RTand the mixture was stirred for 15 min.2-(6-iodohexyl)isoindoline-1,3-dione (2.2 g, 6.24 mmol, 2 eq) was thenadded to the mixture and the resultant mixture was heated at 80° C. for2 h. The reaction was monitored by TLC. After completion, ice-cold water(50 mL) was added to the mixture to obtain a precipitate which wasfiltered over Buchner funnel and dried under vacuum to afford the titlecompound. LC-MS 486[M+H]⁺

Step-5: Preparation of 2-(6-(4-ethyl-6-(2-methyl-6-(1-(tetrahydro-2Hpyran-2-yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)yl)hexyl)isoindoline-1,3-dione

To a stirred solution of2-(6-(6-bromo-4-ethyl-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)isoindoline-1,3-dione(1.0 g, 2.06 mmol) in DMF (20 mL) was added2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(1.52 g, 4.12 mmol, 2.0 eq) and K2CO₃ (0.567 mg, 4.11 mmol, 2 eq)dissolved in water (3 mL) and the mixture was degassed under nitrogenfor 15 min. Pd(dppf)Cl₂ (0.032 mg, 0.043 mmol, 0.1 eq) was then added tothe mixture and the resultant mixture was heated at 70° C. for 16 h. Thereaction monitored by TLC. After completion, the mixture was dilutedwith water (200 mL) and extracted with 10% MeOH/DCM (300 mL×3). Thecombined organic layers were washed with water (100 mL), brine (150 mL)dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford a crude residue which was purified by CombiFlashchromatography to afford the title compound. LC-MS 650 [M+H]⁺

Step-6: Preparation of4-(6-aminohexyl)-1-ethyl-7-(2-methyl-6-(1-(tetrahydro-211 pyran-2yl)-1H-1,2,4-triazol-3 yl)pyridin-3yl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one

To a stirred solution of2-(6-(4-ethyl-6-(2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4triazol-3-yl)pyridin-3-yl)-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)isoindoline-1,3-dione(0.30 g, 1.38 mmol) in EtOH (8 mL) was added NH₂NH₂.H₂O (0.067. 1.38mmol) and the mixture was heated at 100° C. for 16 h. The reactionmonitored by TLC and LC-MS. After completion, the mixture was dilutedwith water (200 mL) and extracted with 10% MeOH/DCM (300 mL×3). Thecombined organic layers were washed with water (100 mL), brine (1.50 mL)dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford the title compound. LC-MS 520[M+H]⁺

Step-7: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-4-hydroxy-5,5-dimethyl-2-thioxoimidazolidin-1yl)-N-(6-(4-ethyl-6-(2-methyl-6-(1-(tetrahydro-2H-pyran-2yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)-2-fluorobenzamide

To a stirred solution of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoicacid (0.08 g, 0.177 mmol) in DMA (6 mL) was added HBTU (0.08 g, 0.212mmol, 1.2 eq) at 0° C. and the resulting mixture was stirred for 10 min.DIPEA (0.067 mL, 0.389 mmol, 2.2 eq) and2-(6-(4-ethyl-6-(2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)isoindoline-1,3-dione(0.110 g, 0.212 mmol, 1.2 eq) were then successively added to themixture and the mixture was stirred at RT for 2 h. The reaction wasmonitored by TLC & LC-MS. After completion, water (10 mL) was added andthe resulting precipitate was filtered over Buchner funnel. The productobtained was washed with water (5 mL×2) and n-pentane (5 mL×2), driedunder vacuum to obtain a crude residue which was purified CombiFlashchromatography to afford the title compound. LC-MS 955 [M+H]⁺

Step-8: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-N-(6-(4-ethyl-6-(2-methyl-6-(1H-1,2,4-triazol-3-yl)pyridin-3-yl)-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)-2-fluorobenzamide

To4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-N-(6-(4-ethyl-6-(2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-3-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)-2-fluorobenzamide(0.18 g, 0.188 mmol) was added 2M-HCl in MeOH (7 mL) and resultingreaction mixture was stirred at RT for 16 h. The reaction was monitoredby TLC & LC-MS. After completion, the mixture was quenched by NaHCO₃solution (30 mL) and extracted with 10% MeOH/DCM (200 mL×2). Thecombined organic layers were washed with water (100 mL), brine (150 mL)dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford a crude residue which was purified by reversed phaseHPLC to afford the title compound. LC-MS 871 [M+H]⁺. ¹HNMR (400 MHz,MeOD-d₄) δ 8.31 (s, 1H), 8.19-8.09 (m, 2H), 8.04-7.95 (m, 3H), 7.93 (s,1H), 7.83 (t, J=8.1 Hz, 1H), 7.39-7.29 (m, 2H), 4.31 (s, 2H), 4.19 (q,J=7.0 Hz, 2H), 3.63 (t, J=7.5 Hz, 2H), 3.43 (t, J=6.9 Hz, 2H), 2.76 (s,3H), 1.93 (d, J=7.4 Hz, 1H), 1.79-1.60 (m, 3H), 1.57 (s, 6H), 1.50 (qd,J=13.4, 10.6, 5.3 Hz, 4H), 1.27 (q, J=7.0, 5.6 Hz, 3H).

Example S-35. Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(5-(7-(2-methyl-6-(1H-1,2,4-triazol-3-yl)pyridin-3-yl)-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)pentyl)benzamide(Compound 3.3)

Step-1: Preparation of ethyl 2-(3,5-dibromopyrazin-2 ylamino)acetate

To a stirred solution of 3,5-dibromopyrazin-2-amine (20.0 g, 79.6 mmol)in DMF (100 mL) was added Cs₂CO₃ (36.2 g, 102.8 mmol) at 0° C. and themixture was stirred for 15 min. Ethyl 2-bromoacetate (10.4 g, 94.8 mmol)was then added to the mixture and the mixture was heated at 80° C. for 3h. The reaction was monitored by TLC. Upon completion, the mixture wasdiluted with water (200 mL) to obtain a precipitate which was filteredover Buchner funnel and dried under vacuum to afford the title compound.LC-MS 338 [M+H]⁺

Step-2′: Preparation of 2-(6-aminohexyl)isoindoline-1,3-dione

To compound 2-(6-iodohexyl)isoindoline-1,3-dione (6.2 g, 17.35 mmol) wasadded liquor NH₃ (35 mL) and the mixture was heated at 80° C. for 3 h.The reaction was monitored by TLC and LC-MS. After completion, themixture was concentrated under reduced pressure and washed withDCM/Hexane (30 mL/100 mL) and dried under vacuum to afford the titlecompound. LC-MS 247 [M+H]⁺

Step-2: Preparation of ethyl2-(5-bromo-3-(5-(1,3-dioxoisoindolin-2-yl)pentylamino)pyrazin-2-ylamino)acetate

To a stirred solution of 2-(5-aminopentyl)isoindoline-1,3-dione (3.2 g,9.44 mmol) in NMP (15 mL) was added DIPEA (8.2 mL, 47.20 mmol, 5 eq) and2-(6-aminohexyl)isoindoline-1,3-dione (4.6 g, 18.88 mmol, 2.0 eq) andthe mixture was heated at 120° C. in sealed tube for 16 h. The reactionwas monitored by TLC and LC-MS. After completion, the mixture wasdiluted with saturated NaHCO₃ solution (50 mL) and extracted with EtOAc(100 mL×2). The combined organic layers were washed with brine (50 mL),dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to obtain a crude which was purified by column chromatographyto afford the title compound. LC-MS 490 [M+H]⁺

Step-3: Preparation of2-(6-(7-bromo-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H) yl)hexyl)isoindoline-1,3-dione

To a stirred solution of ethyl2-(5-bromo-3-(5-(1,3-dioxoisoindolin-2-yl)pentylamino)pyrazin-2-ylamino)acetate (2.0 g, 3.97 mmol) in methanol (5 mL) wasadded AcOH (15 mL) and the mixture was heated at 120° C. in sealed tubefor 16 h. The reaction was monitored by TLC and LC-MS. After completion,the mixture was diluted with saturated NaHCO₃ solution (50 mL) andextracted with EtOAc (100 mL×2). The combined organic layers were washedwith brine (50 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to obtain a crude which was purifiedby column chromatography to afford the title compound. LC-MS 458 [M+H]⁺

Step-4: Preparation of 2-(6-(7-(2-methyl-6-(1-(tetrahydro-2H-pyran-2yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)yl)hexyl)isoindoline-1,3-dione

To a stirred solution of2-(6-(7-bromo-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)isoindoline-1,3-dione (0.20 g, 0.43 mmol) in DMF (6 mL) was added2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(0.32 mg, 0.872 mmol, 2.0 eq) and K2CO₃ (0.12 mg, 0.872 mmol, 2 eq)dissolved in water (3 mL) and the mixture was degassed under nitrogenfor 15 min. Pd(dppf)Cl₂ (0.032 mg, 0.043 mmol, 0.1 eq) was then added tothe mixture and the resultant mixture was heated at 70° C. for 16 h. Thereaction monitored by TLC. After completion, the mixture was dilutedwith water (200 mL) and extracted with DCM (200 mL×3). The combinedorganic layers were washed with water (100 mL), brine (150 mL) driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto afford a crude which was purified by CombiFlash chromatography toafford the title compound. LC-MS 622 [M+H]⁺

Step-5: Preparation of5-(5-amino-2-(p-tolyloxy)phenyl)-1-methyl-3-(methylamino)pyridin-2(1H)-one

To a stirred solution of2-(6-(7-(2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)isoindoline-1,3-dione(0.21 g, 0.427 mmol) in EtOH (5 mL) was added NH₂NH₂.H₂O (0.80 mL) at RTand the mixture was heated at 100° C. for 16 h. The reaction wasmonitored by TLC and LC-MS. After completion, the mixture was dilutedwith water (200 mL) and extracted with 10% MeOH/DCM (200 mL×2). Thecombined organic layers were washed with water (100 mL), brine (150 mL)dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford the title compound. LC-MS 492 [M+H]⁺

Step-6: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(5-(7-(2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)yl)pentyl)benzamide

To a stirred solution of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoicacid (0.12 g, 0.265 mmol) in DMA (7 mL) was added HBTU (0.129 g, 0.319mmol, 1.2 eq) at 0° C. and the mixture was stirred AT 0° C. for 10 min.DIPEA (0.94 mL, 0.524 mmol, 2.2 eq) and5-(5-amino-2-(p-tolyloxy)phenyl)-1-methyl-3-(methylamino)pyridin-2(1H)-one(0.156 g, 0.319 mmol, 1.2 eq) were then successively added to themixture and the mixture was stirred at RT for 2 h. The reaction wasmonitored by TLC & LC-MS. After completion, water (10 mL) was added andthe resulting precipitate was filtered over Buchner funnel. The productobtained was washed with water (5 mL×2) and n-pentane (5 mL×2), driedunder vacuum to obtain a crude product which was purified CombiFlashchromatography to afford the title compound. LC-MS 925 [M+H]⁺

Step-7: Preparation of4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1yl)-2-fluoro-N-(6-(7-(2-methyl-6-(1H-1,2,4-triazol-3-yl)pyridin-3-yl)-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)hexyl)benzamide

The4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-(5-(7-(2-methyl-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazol-3-yl)pyridin-3-yl)-2-oxo-3,4-dihydropyrazino[2,3-b]pyrazin-1(2H)-yl)pentyl)benzamide(0.13 g, 0.140 mmol) was added 2M-HCl in MeOH (10 mL) and resultingreaction mixture was stirred at RT for 16 h. The reaction was monitoredby TLC & LC-MS. After completion, the reaction mixture was quenched byNaHCO₃ solution (25 mL) and extracted with 10% MeOH×DCM (100 mL×2). Thecombined organic layers were washed with water (100 mL), brine (150 mL)dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford a crude residue which was purified by reversed phaseHPLC to afford the title compound. LC-MS 841 [M+H]⁺. ¹H NMR (400 MHz,MeOD-d₄) δ 8.20-8.13 (m, 2H), 8.01 (dd, J=10.3, 7.6 Hz, 4H), 7.85 (s,1H), 7.79 (t, J=8.1 Hz, 1H), 7.38-7.28 (m, 2H), 4.29 (s, 2H), 4.15 (t,J=7.6 Hz, 2H), 3.41-3.34 (m, 2H), 2.76 (s, 3H), 1.76 (q, J=7.3 Hz, 2H),1.67-1.59 (m, 2H), 1.57 (s, 6H), 1.51-1.43 (m, 2H), 1.30 (d, J=8.8 Hz,2H).

The compounds of Table 1 are prepared according to the experimentaldetails exemplified in the Synthetic Examples using the appropriatestarting materials and reagents.

Biological Assays

The following methods are for evaluating the in vitro biology propertiesof the test articles.

a. RBC HotSpot Poly(ADP-ribose) Polymerases 1 (PARP1) andPoly(ADP-ribose) Polymerases 2 (PARP2) Assays: The assay principle isradioisotope-based filter binding assay where incorporation ofradioisotope-labeled NAD⁺ into the substrate captured on filter isdetected after washout free NAD⁺. Data were analyzed using Excel andGraphPad Prism software for IC₅₀ curve fits. Each assay was conductedwith PJ34 as a positive control.

-   -   i. PARP1 assay: Human recombinant PARP1 at a final concentration        of 2.5 nM was combined with histone H4 (20 μM) and test articles        at various concentrations in reaction buffer (50 mM Tris-HCL, pH        8.0, 50 mM NaCl, 10 mM MgCl₂, 1 mM DTT, 1% DMSO, and 20 μg/mL        activated DNA). The solution was inoculated for 20 min at room        temperature and the reaction initiated by adding        [adenylate-³²P]-Nicotinamide Adenine Dinucleotide, ³²P-NAD⁺ at a        final concentration of 10 μM. After incubation for 2 hour at        room temperature, the reaction mixture was filtered and washed        with 0.75% phosphoric acid for radioactivity detection.    -   ii. PARP2 assay: Human recombinant PARP2 at a final        concentration of 2.5 nM was combined with histone H3 (20 μM) and        test articles at various concentrations in reaction buffer (50        mM Tris-HCL, pH 8.0, 50 mM NaCl, 10 mM MgCl₂, 1 mM DTT, 1% DMSO,        and 20 μg/mL activated DNA). The solution was inoculated for 20        min at room temperature and the reaction was initiated by adding        [adenylate-³²P]-Nicotinamide Adenine Dinucleotide, ³²P-NAD⁺ at a        final concentration of 10 μM. After incubation for 2 hour at        room temperature, the reaction mixture was filtered and washed        with 0.75% phosphoric acid for radioactivity detection.        b. AR binding assay: AR in LNCaP cytosol was used for        determining the binding affinity of test articles and the        reference compound—progesterone (Sigma, Cat: E2785, St. Louis,        Mo.). IC₅₀s were determined using 8 concentrations/compound.        Cytosol was plated at 200 μg/well (100 μL) into a 96-well        conical polypropylene plate (Agilent, Cat: 5042-1385, Santa        Clara, Calif.) and mixed with 3 μL of test compound. After        adding 100 μL of ³H-methyltrienolone (PerkinElmer, Cat:        NET590250UC, San Jose, Calif.), the plate was sealed and shaken        at 300 rpm at 4° C. for 24 hours. Post incubation, 100 μL of        radioligand adsorption buffer containing 10 mM Tris-HCl, pH 7.4;        1.5 mM EDTA; 1 mM DTT; 0.25% charcoal; 0.0025% dextran was added        to individual well. Plate was shaken for 15 min at 4′C followed        by centrifugation at 3000 rpm for 30 min at 4° C. 150 μL of        supernatant was transferred into scint-tube (PerkinElmer,        Cat: 6000192) and mixed with 2 mL of Ultima Gold Cocktail        (PerkinElmer, Cat: 6013329). Radioactivity was counted using a        TriCarb 2910 TR scintillation counter (PerkinElmer). Inhibition        of the radioactivity by test articles were calculated using the        equation below:

% Inhibition=(1−(Assay well−Average_LC)/(Average_HC−Average_LC))×100%.

IC₅₀ values were calculated and graphed using the model “log(inhibitor)vs. response—Variable slope” included in GraphPad Prism 5 (San Diego,Calif.). The Ki values were further calculated using the equation belowwhere [L] was the radioligand concentration (1 nM) using in this study.Kd value was 0.332 nM. Results are listed in Table 3.

Ki=IC₅₀/(1+[L]/Kd)

c. AR transactivation: Human AR cloned into a CMV vector backbone wasused for the transactivation study. HEK-293 cells were plated at 80,000cells per well of a 24 well plate in DME+5% csFBS. Twenty four hourslater, the cells were transfected using Lipofectamine (Invitrogen,Carlsbad, Calif.) with 0.25 μg GRE-LUC, 0.01 μg CMV-LUC (renillaluciferase) and 25 ng of the AR in OPTIMEM medium. The cells weretreated 24 hrs after transfection with various ligands (10⁻¹² to 10⁻⁵ Mfinal concentrations) and luciferase assay were performed 48 hours aftertransfection. Firefly values were normalized to renilla luciferasevalues and the values were represented as relative light units (RLU).Agonist and antagonist assays for the test article were performed in theabsence and in the combination with 0.1 nM R1881, respectively. Datawere represented as EC₅₀ (for agonists) and IC₅₀ (for antagonists)obtained from four parameter logistics curve. Each experiment wasperformed with R1881 as an agonist Results for AR antagonism are shownin Table 2.d. Cell culture and proliferation assays: LNCaP, PC-3, 22RV1, MCF-7,HT-29, and HCT-116 cells were procured from American Type CultureCollection (ATCC). Cells were cultured in medium recommended by theATCC. Cell culture medium was obtained from Fisher scientific (Waltham,Mass.) and serum was obtained from Hyclone (San Angelo, Tex.).

Cells were plated at varying density in the respective growth medium in96 well plates. 24 hr later, cells were treated, in triplicate orquadruplicate, with test articles prepared in a range of concentrationsby series dilution of DMSO stock solutions in growth medium andincubated for three to seven days. The number of viable 22RV1 and HT-29cells was measured using CellTiter Glo assay (CTG, Promega, Madison,Wis.) after three days of treatment. The other types of cancer cellswere re-treated at three or four days and the number of viable cells wasmeasured using CTG assay after a total treatment time of six or sevendays. Cell viability data were plotted using GraphPad Prism (GraphPadSoftware, Inc., San Diego, Calif.). In addition, a nonlinear regressionmodel with a sigmoidal dose response and variable slope within GraphPadPrism was used to calculate the IC₅₀ value of individual test articles.Results of the assays are shown in Table 4.

e. Additional cell proliferation assays: Other cancer cell lines aretested in a cell proliferation assay. For example, SK-OV-3 or OVCAR3ovarian cancer cells are tested. Cells are cultured in mediumrecommended by the supplier (e.g., ATCC or JCRB Cell Bank), at 37° C. ina 5% CO₂ environment. For the proliferation assay, cells are plated inthe growth medium in 96 well plates. Seeding density is adjustedaccording to the cell type. 24 hr later, cells are treated, intriplicate or quadruplicate, with test articles prepared in a range ofconcentrations by series dilution of DMSO stock solutions in growthmedium, and typically incubated for three to seven days, with testarticle-containing medium replaced after three or four days. The numberof viable cells is measured using CellTiter Glo assay (CTG, Promega,Madison, Wis.) or similar. Cell viability data are plotted usingGraphPad Prism (GraphPad Software, Inc., San Diego, Calif.). Inaddition, a nonlinear regression model with a sigmoidal dose responseand variable slope within GraphPad Prism is used to calculate the IC₅₀value of individual test articles. Similarly, HEK-293 and HeLa calls canalso be tested by methods known in the art.f. Nuclear translocation: LNCaP cells are plated on coverslips in 24well plates in growth medium. Twenty-four hours after plating, medium ischanged to RPMI+1% csFBS and the cells are maintained in this medium fortwo days. Medium is replaced again and the cells are treated. Cells arefixed 4 hours after treatment and the AR immunostained using AR N20antibody (Santa Cruz Biotechnology, Santa Cruz, Calif.). Nucleus isstained with DAPI. Cells are imaged with a confocal microscope.g. ER binding assay: ERα binding was assessed by the LanthaScreen®TR-FRET ER Alpha Competitive Binding Assay at Thermo Fisher. In thisassay, a terbium-labeled anti-GST antibody was used to indirectly labelGST-tagged ER Alpha-ligand binding domain (LBD) by binding to its GSTtag. Competitive binding to the ER Alpha-LBD (GST) was detected by atest compound's ability to displace a fluorescent ligand (Fluormone™ ES2Green tracer) from the ER Alpha-LBD (GST), which results in a loss ofFRET signal between the Tb-anti-GST antibody and the tracer. Whenrunning the assay, Fluormone™ ES2 Green tracer was added to ligand testcompounds or solvent controls followed by addition of a mixture of theER Alpha-LBD (GST) and terbium anti-GST antibody. After an incubationperiod at room temperature, the TR-FRET ratio of 520:495 emissions werecalculated and was used to determine the IC₅₀ from a dose response curveof the compound. Results of the assay are shown in Table 3.h. ER and PR functional assays: COS cells are transfected with 25 ng ratprogesterone receptor (PR) and 250 ng GRE-LUC or 50 ng human estrogenreceptor a (ER) and 250 ng ERE-LUC. Cells are also transfected with 10ng CMV-renilla LUC in OptiMEM medium using lipofectamine transfectionreagent. Twenty-our hours after transfection medium is changed to DME+5%csFBS w/o and treated with compounds in the presence of 0.1 nMprogesterone for PR and estradiol for ER. Twenty four hours aftertreatment, cells are harvested and luciferase assay is performed usingdual luciferase assay kit. The firefly values are normalized to renillaluciferase values and represented as a ratio.i. Evaluation of test compound in mouse xenograft model: To examine thein vivo antitumor activity of test compound, tumor growth experimentsare performed in a cell line xenograft model. Male NOD SCID Gamma (NSG)mice are housed as five animals per cage and are allowed free access towater and commercial rodent chow. 22RV1 cells (grown in RPM1+10% FBS)mixed with 50% matrigel basement membrane are implanted subcutaneouslyin castrated mice. Alternatively, an antiandrogen resistant cell lineother than 22RV1, such as MR49F or VCaP, is used. Once the tumors reach200-500 mm³, the animals are randomized and treated intraperitoneallywith vehicle (DMSO:PEG-300:corn oil 10:30:60 ratio) or test compound.Tumors are measured thrice weekly and the volume is calculated using theformula length*width*width*0.5. Animals are sacrificed at the end of 28days of treatment and the tumors are weighed and stored for furtherprocessing. The tumor growth inhibition (TGI) is calculated by comparingthe control group's tumor measurements with the other study groups. TGIis calculated for each group using the formula listed below:

TGI(%)=[1−(TV_(Treatment_DayN)−TV_(Treatment_Day0))/(TV_(Vehicle_DayN)−TV_(Vehicle_Day0))]×100%

TV_(Treatment_DayN) is the average tumor volume of a treatment group ona given day, TV_(Treatment_Day0) is the average tumor volume of thetreatment group on the first day of treatment, TV_(Vehicle_DayN) is theaverage tumor volume of the vehicle control group on a given day, andTV_(Vehicle_Day0) is the average tumor volume of the vehicle group onthe first day of treatment.j. DNA-PK Inhibition Inhibition of DNA-PK activity by test compounds wasassessed in a radioisotope-based filter binding assay (Reaction BiologyCorporation HotSpot Kinase Assay). Test compounds were dissolved in 100%DMSO to specific concentrations. Serial dilution of the test compoundswas conducted by Integra Viaflo Assist in DMSO. The substrate,DNA-PKtide (Anaspec, Fremont, Calif.; #60210-5), was prepared inReaction Buffer (20 mM Hepes (pH 7.5), 10 mM MgCl₂, 1 mM EGTA, 0.02%Brij35, 0.02 mg/ml BSA, 0.1 mM Na₃VO₄, 2 mM DTT, 1% DMSO) so that itsfinal concentration in the reaction would be 20 μM. DNA-PK activatorcontaining dsDNA was delivered to the solution (10 μg/mL in finalreaction). DNA-PK (Invitrogen, Carlsbad, Calif.; #PR9107A) was deliveredto the substrate solution (5 nM in final reaction) and the solutiongently mixed. Compounds in 100% DMSO were delivered into the kinasereaction mixture by Acoustic technology (Echo550; nanoliter range), andthe reaction incubated for 20 min at room temperature. ³³P-ATP (Specificactivity 10 μCi/μL) was delivered into the reaction mixture to initiatethe reaction, which was incubated for 2 hours at room temperature.Radioactivity was detected by a filter-binding method. Kinase activitydata were expressed as the percent remaining kinase activity in testsamples compared to vehicle (dimethyl sulfoxide) reactions. IC₅₀ valuesand curve fits were obtained using GraphPad Prism. Results of the assayare shown in Table 5.

TABLE 2 In vitro results Compound PARP1 PARP2 AR Ant. No. IC₅₀ (μM) IC₅₀(μM) IC₅₀ (μM) 1.1 0.002 0.0002 0.033 1.2 0.003 0.0002 0.481 1.3 0.0020.0005 ent-1.3 0.079 0.016 1.4 0.002 0.0004 1.5 0.023 0.002 0.75 1.680.0003 0.0002 3.3 1.83 0.0007 0.0003 0.57 1.96 0.006 0.0005 1.97 0.0020.0004 1.98 0.0002 0.0001 1.99 0.0008 0.0002 1.104 0.0012 0.0003 1.1070.0003 0.0001 1.110 0.0007 0.0002 1.111 0.0016 0.0002 1.114 0.02790.0019 1.115 0.0257 0.0015 1.116 0.0005 0.0002 1.117 0.0003 0.0001 1.1180.0136 0.0010 1.119 0.0016 0.0003 2.4 0.004 0.0003 2.13 0.0033 0.00052.14 0.274 0.0124

TABLE 3 In vitro NHR binding Compound No. AR K_(i) (nM) ERαK_(i) (nM)1.2 1.31 1.5 291 1.68 907 1.83 139 1.97 504 1.98 181 1.99 197 1.104 1671.107 >7469 1.110 60 1.111 36 1.114 19 1.115 30 1.116 4969 1.117 601.118 >7469 1.119 20 2.4 5 2.13 3.3 2.14 3.5 3.1 21 3.2 19 3.3 14

TABLE 4 In-vitro cell growth inhibition 22RV1 MCF-7 HT-29 LNCaP PC-3HCT-116 Compound IC₅₀ IC₅₀ IC₅₀ IC₅₀ IC₅₀ IC₅₀ No. (μM) (μM) (μM) (μM)(μM) (μM) 1.1 0.30 0.59 0.05 0.31 1.2 0.40 >10 0.79 0.24 0.11 1.3 0.498.8 >30 ent-1.3 >100 1.4 0.09 0.02 0.11 1.5 4.5 5.0 1.68 4.7 >101.83 >30 >10 1.97 >30 >30 1.98 3.5 1.99 3.6 1.104 1.1 9.3 1.107 1.31.110 0.84 9.5 1.111 0.89 1.114 1.3 1.115 3.5 1.116 2.6 1.117 0.44 1.1189.7 1.119 2.9 2.2 0.39 2.4 11 2.13 1.69 3.6 6.0 2.14 9.3 >30

TABLE 5 In vitro enzyme inhibition Compound DNA-PK No. IC₅₀ (μM) 3.1 >103.2 >10 3.3 0.0018

1. A compound comprising at least one nuclear payload and at least onenuclear receptor-targeting epitope; provided that when the compoundcomprises one nuclear payload and one nuclear receptor-targetingepitope: the nuclear receptor-targeting epitope is not a peptide,protein, nanoparticle or antibody; and when the nuclearreceptor-targeting epitope is an androgen receptor-targeting epitope oran estrogen receptor-targeting epitope, the nuclear payload is notdoxorubicin, or an analog thereof, and is not a hydroxamic acid whichbinds histone deacetylase (HDAC).
 2. A compound comprising at least onenuclear payload which binds to a catalytic domain of poly(ADP-ribose)polymerase (PARP) and at least one nuclear receptor-targeting epitope.3. The compound of claim 1 or 2, wherein at least one nuclearreceptor-targeting epitope is a nuclear steroid receptor-targetingepitope.
 4. The compound of any preceding claim, wherein the nuclearpayload is bonded to one nuclear receptor-targeting epitope via anon-biocleavable linking moiety and one or more nuclearreceptor-targeting epitope(s) via a biocleavable linking moiety.
 5. Thecompound of any preceding claim, wherein the compound comprises at leastone nuclear payload which binds to a protein involved in the DNA damagerepair process.
 6. The compound of any preceding claim, wherein thecompound comprises at least one nuclear payload which bindspoly(ADP-ribose) polymerase (PARP), DNA-dependent protein kinase(DNA-PK), histone deacetylase (HDAC), enhancer of zeste homolog 2(EZH2), histone acetyl transferase (HAT), methyltransferase, abromodomain, myelin transcription factor 1 (MYT1), p53,melanocyte-stimulating hormone (MSH), mutL homolog (MLH), ERCC1,apurinic/apyrimidinic endonuclease 1 (APE1), topoisomerase I (Topo I),topoisomerase II (Topo II), Wee1, checkpoint kinase1 (Chk1), checkpointkinase2 (Chk2), ataxia telangiectasia (ATR), or ataxia-telangiectasiamutated (ATM).
 7. The compound of any preceding claim, wherein thenuclear payload comprises olaparib (AZD-2281), Olaparib TOPARP-A,rucaparib (AG014699, PF-01367338), niraparib, talazoparib (BMN-673),veliparib (ABT-888), CEP 9722, E7016, BGB-290, 3-aminobenzamide,methoxyamine, CC-11.5, MSC2490484A, AZD6738, VX-970, AZD0156, GDC-0.575,MK-8776, LY2606368, AZD1775, belotecan, CRLX101, irinotecan, LMP 400,LMP 776, NKTR-102, topotecan, doxorubicin, epirubicin, etoposide,idarubicin, mitoxantrone, teniposide, vorinostat, romidepsin (Istodax),chidamide, panobinostat (Farydak), belinostat (PXD101), panobinostat(LBH589), valproic acid (as Mg valproate), mocctinostat (MGCD0103),abcxinostat (PCI-24781), entinostat (MS-275), SB939, resminostat(4SC-201), givinostat (ITF2357), quisinostat (JNJ-26481585), HBI-8000,kevetrin, CUDC-101, AR-42, CHR-2845, CHR-3996, 4SC-202, CG200745,ACY-1215, ME-344, sulforaphane, tazemetostat, MAK638, CPI-1205),DS-3201b, anacardic acid, MG149, C646, S-adenosyl methionine, JQ1, I-BET151 (GSK1210151A), I-BET 762 (GSK525762), OTX-015, TEN-010, CPI-203,CPI-0610, olinone, LY294002, or an analog thereof.
 8. The compound ofclaim 2, wherein the compound comprises at least one nuclear payloadwhich binds to a catalytic domain of poly(ADP-ribose) polymerase (PARP).9. The compound of claim 2, wherein the catalytic domain comprises aconserved HYE motif.
 10. The compound of any preceding claim, whereinthe compound comprises at least one nuclear payload which binds to apoly(ADP-ribose) polymerase (PARP) with an IC₅₀ of less than about 500nM.
 11. The compound of claim 8, wherein the PARP comprises PARP-1and/or PARP-2, or a variant thereof.
 12. The compound of any precedingclaim, wherein the compound is of Formula I or II:A-(L-B)_(m)  IA-L-(B)_(m)  II wherein: A is a nuclear payload; m is 1, 2 or 3; each Bis independently a nuclear receptor-targeting epitope; and each L isindependently a covalent bond or a linking moiety.
 13. A compound ofFormula III, IV, V or VI, or stereoisomer, mixture of stereoisomers,hydrate, solvate, isotopically enriched analog or pharmaceuticallyacceptable salt thereof:

wherein: each R¹, R², R³ and R⁴ is independently -L-(B)_(m), hydrogen,C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₀ cycloalkyl,heterocyclyl, aryl, heteroaryl, —C(═O)R⁵, —C(═O)OR⁵, —C(═O)NR⁵R⁶,—S(═O)₁₋₂R⁵, —S(═O)₁₋₂NR⁵R⁶, —NR⁵S(═O)₁₋₂R⁶ or —C═NOR⁵, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroarylof R¹, R², R³ and R⁴ are independently optionally substituted with oneor more R¹⁰ as valency permits; m is 1, 2, or 3; each L is independentlya covalent bond or a linking moiety; each B is independently a nuclearreceptor-targeting epitope; each R¹⁰ is independently halo, cyano,nitro, —OR⁷, —SR⁷, —SF₅, —NR⁷R⁸, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂alkynyl, C₃₋₁₀ cycloalkyl, heterocyclyl, aryl, heteroaryl, —C(═O)R⁷,—C(═O)OR⁷, —OC(═O)OR⁷, —OC(═O)R⁷, —C(═O)NR⁷R⁸, —OC(═O)NR⁷R⁸,—NR⁷C(═O)NR⁷R⁸, —S(═O)₁₋₂R⁷, —S(═O)₁₋₂NR⁷R⁸, —NR⁷S(═O)₁₋₂R⁸,—NR⁷S(═O)₁₋₂NR⁷R⁸, —NR⁷C(═O)R⁸, —NR⁷C(═O)OR⁸ or —C═NOR⁷, wherein eachalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, awl and heteroaryl ofR¹⁰ are independently optionally substituted with one or more halo orC₁₋₁₂ alkyl optionally substituted by oxo, halo, hydroxyl or amino asvalency permits; and each R⁵ and R⁶ is independently hydrogen,deuterium, C₁₋₁₂ alkyl or C₃₋₁₂ cycloalkyl, optionally substituted withoxo, halo, hydroxyl or amino as valency permits; or R⁵ and R⁶ are takentogether with the atoms to which they are attached to form heterocyclyloptionally substituted by halo or C₁₋₁₂ alkyl optionally substituted byoxo, halo, hydroxyl or amino; each R⁷ and R⁸ is independently hydrogen,deuterium or C₁₋₁₂ alkyl optionally substituted with oxo, halo, hydroxylor amino as valency permits; or R⁷ and R⁸ are taken together with theatoms to which they are attached to form heterocyclyl optionallysubstituted by halo or C₁₋₁₂ alkyl optionally substituted by oxo, halo,hydroxyl or amino; and R⁹ is hydrogen or R²; provided that at least oneR¹, R², R³ and R⁴ is -L-(B)_(m).
 14. A compound of Formula IIIB, IVA, VBor VIA, or stereoisomer, mixture of stereoisomers, hydrate, solvate,isotopically enriched analog or pharmaceutically acceptable saltthereof:

wherein: R² is -L-(B)_(m); m is 1, 2, or 3; L is a covalent bond or alinking moiety; and each B is independently a nuclear receptor-targetingepitope.
 15. The compound of any one of claims 12-14, wherein at leastone L is a non-biocleavable linking moiety.
 16. The compound of any oneof claims 12-14, wherein at least one L is an acid-labile linkingmoiety.
 17. The compound of any one of claims 12-16, wherein the linkingmoiety is alkylene, heteroalkylene, alkenylene, heteroalkenylene,alkynylene, heteroalkynylene, arylene, heteroarylene, cycloalkylene orheterocycloalkylene; wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, may optionally comprisean arylene, heteroarylene, cycloalkylene or heterocycloalkylene; andfurther wherein each alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, arylene, heteroarylene,cycloalkylene or heterocycloalkylene is independently optionallysubstituted with one to five substituents independently selected fromoxo, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkyl.
 18. The compoundof any one of claims 12-17, wherein the linking moiety is of theformula:—Y¹—(CH₂)_(n′)—Y²—(CH₂)_(m′)—Y³— wherein: each of Y¹, Y², and Y³ areindependently a bond, —NR¹¹—, —O—, —S(O)₀₋₂—, —NR¹¹C(O)—, C(O)NR¹¹—,—NR¹¹S(O)₂—, —S(O)₂NR¹¹—, —CR¹²═N—NR¹¹—, —NR¹¹—N═CR¹²—, —C(O)—, arylene,heteroarylene, cycloalkylene or heterocycloalkylene; wherein eachalkylene, heteroalkylene, alkenylene, heteroalkenylene, alkynylene,heteroalkynylene, arylene, heteroarylene, cycloalkylene orheterocycloalkylene is independently optionally substituted with one tofive substituents independently selected from oxo, halo, C₁₋₄ alkyl,C₁₋₄ alkoxy, and C₁₋₄ haloalkyl; each R¹¹ is independently C₁₋₄ alkyl,C₁₋₄ haloalkyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; each R¹²is independently C₁₋₄ alkyl, C₁₋₄ haloalkyl, aryl, heteroaryl,cycloalkyl or heterocyclyl; and n′ and m′ are each independently 0, 1,2, 3, 4, 5, 6, 7, or
 8. 19. The compound of any one of claims 12-18,wherein at least one linking moiety comprises a hydrazone linkage. 20.The compound of any preceding claim, wherein the at least one nuclearreceptor-targeting epitope is a nuclear steroid receptor-targetingepitope.
 21. The compound of any preceding claim, wherein the at leastone nuclear receptor-targeting epitope is independently selected from anestrogen receptor-targeting epitope, glucocorticoid receptor-targetingepitope, progesterone receptor-targeting epitope or androgenreceptor-targeting epitope.
 22. The compound of any preceding claim,wherein the at least one nuclear receptor-targeting epitopeindependently comprises an epitope derived from an androgen receptoragonist, an androgen receptor antagonist, a selective androgen-receptormodulator (SARM), an estrogen receptor agonist, an estrogen receptorantagonist, a selective estrogen receptor modulator (SERM), aglucocorticoid receptor antagonist, a glucocorticoid receptor agonist, aselective glucocorticoid receptor modulator (SGRM), a progesteronereceptor antagonist, a progesterone receptor agonist, a selectiveprogesterone receptor modulator (SPRM), or a combination thereof. 23.The compound of any preceding claim, wherein the at least one nuclearreceptor-targeting epitope independently comprises an epitope derivedfrom estrogen, estetrol, estriol, estrone, progesterone, enobosarm,bicalutamide, apalutamide, testosterone, dihydrotestosterone, estradiol,flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene,bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone,LGD-2941, BMS-564929, ostarine, or an analog thereof.
 24. The compoundof any preceding claim, wherein at least one nuclear receptor-targetingepitope is capable of binding to a ligand binding domain of a nuclearsteroid receptor.
 25. The compound of claim 24, wherein the nuclearsteroid receptor is an estrogen receptor, a glucocorticoid receptor, aprogesterone receptor, or an androgen receptor.
 26. The compound of anypreceding claim, wherein the at least one nuclear receptor-targetingepitope binds to a nuclear steroid receptor with an IC₅₀ of less thanabout 500 nM or an EC₅₀ of less than about 1 μM.
 27. The compound of anypreceding claim, wherein the at least one nuclear receptor-targetingepitope is not a peptide, protein, nanoparticle or antibody.
 28. Acompound as provided in Table
 1. 29. A pharmaceutical compositioncomprising a compound as in any preceding claim and a pharmaceuticallyacceptable excipient.
 30. A method for the treatment or prevention of acondition which can be ameliorated by inhibition of PARP in anindividual in need thereof, the method comprising administering to theindividual an effective amount of a compound or pharmaceuticalcomposition of any preceding claim or a pharmaceutically acceptable saltor solvate thereof.
 31. A method of treating or preventing cancer,comprising administering an effective amount of a compound orpharmaceutical composition of any preceding claim or a pharmaceuticallyacceptable salt or solvate thereof to an individual in need thereof. 32.The method of claim 30 or 31, wherein the administering comprises oraladministration.
 33. The method of any one of claims 30-32, furthercomprising administering an additional chemotherapeutic agent.
 34. Themethod of claim 33, wherein the additional chemotherapeutic agent iscisplatin or etoposide, irinotecan, camptostar, topotecan, paclitaxel,docetaxel, epothilones, taxotere, tamoxifen, 5-fluorouracil,methoxtrexate, temozolomide, cyclophosphamide, SCH 66336, R115777,L778123, BMS 214662, gefitinib, erlotinib hydrochloride, antibodies toEGFR, matinib, intron, cytarabine, adriamycin, cytoxan, gemcitabine,uracil mustard, chlormethine, ifosfamide, melphalan, chlorambucil,pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan,carmustine, lomustine, streptozocin, dacarbazine, floxuridine,cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate,pentostatine, vinblastine, vincristine, vindesine, bleomycin,doxorubicin, dactinomycin, daunorubicin, epirubicin, idarubicin,mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide,17α-ethinylestradiol, diethylstilbestrol, testosterone, prednisone,fluoxymesterone, dromostanolone propionate, testolactone,megestrolacetate, methylprednisolone, methyltestosterone, prednisolone,triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide,estramustine, medroxyprogesterone acetate, leuprolide, flutamide,toremifene, goserelin, carboplatin, hydroxyurea, amsacrine,procarbazine, mitotane, mitoxantrone, levamisol, navelbine, anastrazole,letrazole, capecitabine, reloxafine, droloxifene, hexamethylmelamine,avastin, herceptin, bexxar, velcade, zevalin, trisenox, xeloda,vinorelbine, porfimer, cetuximab, liposomal, Thiotepa, Altretamine,melphalan, Trastuzumab, lerozole, fulvestrant, exemestane, ifosfomide,rituximab, C225, Campath, carboplatin, procarbazine, mechlorethamine,cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil,busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin,bleomycin, plicomycin, mitomycin, etoposide (VP 16), tamoxifen,raloxifene, estrogen receptor binding agents, paclitaxel, gemcitabine,navelbine, farnesyl-protein transferase inhibitors, transplatinum,5-fluorouracil, vincristine, vinblastine and methotrexate, or an analogor derivative thereof.
 35. The method of any one of claims 31-34,further comprising administering radiotherapy to the patient.
 36. Themethod of any one of claims 31-35, wherein the cancer is a BRCA positivecancer.
 37. The method of any one of claims 31-35, wherein the cancer isa solid tumor.
 38. The method of any one of claims 31-35, wherein thecancer is a cancer affecting B cells.
 39. The method of any one ofclaims 31-38, wherein the cancer is a blood cancer, lung cancer, breastcancer, fallopian tube cancer, brain cancer, head and neck cancer,esophageal cancer, ovarian cancer, pancreatic cancer, peritoneal cancer,prostate cancer or skin cancer.
 40. The method of claim 39, wherein thecancer is liver cancer, melanoma, Hodgkin's disease, non-Hodgkin'slymphomas, acute lymphocytic leukemia, chronic lymphocytic leukemia,multiple myeloma, neuroblastoma, breast carcinoma, ovarian carcinoma,lung carcinoma, Wilms' tumor, cervical carcinoma, testicular carcinoma,soft-tissue sarcoma, chronic lymphocytic leukemia, primarymacroglobulinemia, bladder carcinoma, chronic granulocytic leukemia,primary brain carcinoma, malignant melanoma, small-cell lung carcinoma,stomach carcinoma, colon carcinoma, malignant pancreatic insulinoma,malignant carcinoid carcinoma, malignant melanoma, choriocarcinoma,mycosis fungoide, head neck carcinoma, osteogenic sarcoma, pancreaticcarcinoma, acute granulocytic leukemia, hairy cell leukemia,rhabdomyosarcoma, Kaposi's sarcoma, genitourinary carcinoma, thyroidcarcinoma, esophageal carcinoma, malignant hypercalcemia, cervicalhyperplasia, renal cell carcinoma, endometrial carcinoma, polycythemiavera, essential thrombocytosis, adrenal cortex carcinoma, skin cancer,or prostatic carcinoma.
 41. The method of claim 39, wherein the canceris bladder cancer, breast cancer, fallopian tube cancer, ovarian cancer,prostate cancer, peritoneal cancer, testicular cancer, endometrialcancer, or uterine cancer.
 42. A method of treating or preventing anandrogen receptor overexpressing cancer, comprising administering aneffective amount of a compound, or a pharmaceutically acceptable salt orsolvate thereof, comprising at least one nuclear payload and at leastone androgen receptor-targeting epitope to an individual in needthereof.
 43. The method of claim 42, wherein the cancer is prostate,breast, triple negative breast cancer, bladder, or liver cancer.
 44. Themethod of claim 42, wherein the androgen receptor-targeting epitopecomprises an androgen receptor agonist, a selective androgen-receptormodulator (SARM), an androgen receptor antagonist, a selective estrogenreceptor modulator (SERM), an estrogen receptor antagonist, a progestin,or an estrogen.
 45. The method of claim 44, wherein the androgenreceptor-targeting epitope comprises enobosarm, bicalutamide, flutamide,nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene,fulvestrant, megestrol acetate, estramustine, ketoconazole, abiraterone,darolutamide, or an analog thereof.
 46. A method of treating orpreventing an estrogen and/or progesterone receptor overexpressingcancer, comprising administering an effective amount of a compound, or apharmaceutically acceptable salt or solvate thereof, comprising at leastone nuclear payload and at least one estrogen and/or progesteronereceptor-targeting epitope to an individual in need thereof.
 47. Themethod of claim 46, wherein the cancer is breast, uterine, or ovariancancer.
 48. A method of treating or preventing a glucocorticoid receptoroverexpressing cancer, comprising administering an effective amount of acompound, or a pharmaceutically acceptable salt or solvate thereof,comprising at least one nuclear payload and at least one glucocorticoidreceptor-targeting epitope to an individual in need thereof.
 49. Themethod of claim 48, wherein the cancer is prostate, breast, uterine, orovarian cancer.